JP2022534961A - Azepine as a HBV Capsid Assembly Modulator - Google Patents

Abstract

Compounds, compositions and methods for treating diseases, conditions, conditions and disorders affected by modulation of CAM1 are disclosed. Such compounds are represented by the following formula (I): TIFF2022534961000040.tif70170. wherein R1, R2, R3, R4, X and Y are defined herein.

Description

The present disclosure relates to azepine compounds, pharmaceutical compositions containing these compounds, chemical processes for preparing these compounds and their use in the treatment of diseases associated with HBV infection in animals, particularly humans.

Chronic hepatitis B virus (HBV) infection is a major global health problem, affecting over 5% of the world's population (over 350 million people worldwide and 1.25 million individuals in the United States). have an impact on

Despite the availability of a prophylactic HBV vaccine, it is a sub-optimal treatment option, and with new infection rates persisting in most parts of the developing world, the burden of chronic HBV infection is It remains a major unaddressed global health care problem. Current treatments are not curative, are limited to only two classes of drugs (interferon-alpha and nucleoside analogues/inhibitors of viral polymerases), and problems with drug resistance, low efficacy and tolerability limit their effectiveness. The low cure rate of HBV is due, at least in part, to the fact that it is difficult to achieve complete suppression of virus production with a single antiviral agent. However, sustained suppression of HBV DNA is useful for slowing the progression of liver disease and preventing hepatocellular carcinoma. Current therapeutic goals for HBV-infected patients are aimed at reducing serum HBV DNA to low or undetectable levels, ultimately reducing or preventing the development of cirrhosis and hepatocellular carcinoma. there is

HBV capsid proteins perform essential functions in the viral life cycle. The HBV capsid/core protein forms a metastable viral particle or protein shell that protects the viral genome during intracellular transit and is central in viral replication processes such as genome encapsidation, genome replication and virion morphogenesis and release. also play a role. The capsid structure also responds to environmental factors that allow uncoating after viral entry. Consistently, proper timing of capsid assembly and disassembly, proper capsid stability and core protein function have been found to be crucial for viral infectivity.

The crucial function of the HBV capsid protein imposes strict evolutionary constraints on the viral capsid protein sequence, resulting in the observed low sequence variability and high conservation. Consistently, mutations in the HBV capsid that disrupt its assembly are lethal, and mutations that disrupt capsid stability severely attenuate viral replication. A high degree of functional constraint on the multifunctional HBV core/capsid protein is consistent with a high degree of sequence conservation, as many mutations are detrimental to function. In fact, the core/capsid protein sequences are over 90% identical across HBV genotypes, exhibiting only a few polymorphic residues. Therefore, resistance selection against HBV core/capsid protein binding compounds can be difficult to select without significantly affecting viral replication fitness.

Reports describing compounds that bind to the viral capsid and inhibit HIV, rhinovirus and HBV replication provide a strong pharmacological proof of concept for viral capsid proteins as antiviral drug targets. .

There is a need in the art for therapeutic agents that can increase suppression of viral production and that can treat, ameliorate and/or prevent HBV infection. Administration of such therapeutics to HBV-infected patients, either as monotherapy or in combination with other HBV therapies or adjunctive therapies, has been shown to significantly reduce viral load, improve prognosis, reduce disease progression, and reduce serocytosis. It will lead to improvement of conversion rate.

Given the clinical importance of HBV, identifying compounds that can increase suppression of viral production and that can treat, ameliorate and/or prevent HBV infection is attractive for developing new therapeutic agents. is a means. Provided herein are such compounds.

（式中、
Ｒ^１は、Ｆ、ＯＨ及びＣ_１～６アルキルからなる群から選択され、ここで、アルキルは、ＯＨで任意選択的に置換されており；
Ｒ^２は、Ｂｒ、ＣＮ及びＣ_１～４ハロアルキルからなる群から選択され；
Ｒ^３は、Ｈ又はＦであり；
Ｒ^４は、Ｈ又はＣ_１～４アルキルであり；
Ｘは、Ｏ、Ｓ、Ｓ＝Ｏ及びＳＯ_２からなる群から選択され；及び
Ｙは、ＣＨ、ＣＦ及びＮからなる群から選択される）
の化合物及び式（Ｉ）の化合物の薬学的に許容される塩、立体異性体、同位体変異体、Ｎ－オキシド又は溶媒和物を対象とする。 The present disclosure is directed to general and preferred embodiments as defined in independent and dependent claims respectively appended hereto, which are incorporated herein by reference. In particular, the present disclosure provides compounds of formula (I):

(In the formula,
R ¹ is selected from the group consisting of F, OH and C _1-6 alkyl, wherein alkyl is optionally substituted with OH;
R ² is selected from the group consisting of Br, CN and C _1-4 haloalkyl;
R ³ is H or F;
R ⁴ is H or C _1-4 alkyl;
X is selected from the group consisting of O, S, S ₌ O and SO2; and Y is selected from the group consisting of CH, CF and N)
and pharmaceutically acceptable salts, stereoisomers, isotopic variants, N-oxides or solvates of compounds of formula (I).

Further embodiments are pharmaceutically acceptable salts of compounds of formula (I), pharmaceutically acceptable prodrugs of compounds of formula (I), pharmaceutically acceptable metabolisms of compounds of formula (I) products, and enantiomers and diastereomers of the compounds of formula (I), and pharmaceutically acceptable salts thereof.

In embodiments, the compound of formula (I) is a compound selected from those species described or exemplified in the detailed description below.

The present disclosure provides one or more compounds of formula (I), pharmaceutically acceptable salts of compounds of formula (I), pharmaceutically acceptable prodrugs of compounds of formula (I) and compounds of formula (I). Also of interest are pharmaceutical compositions comprising pharmaceutically active metabolites of A pharmaceutical composition may further comprise one or more pharmaceutically acceptable excipients or one or more other agents or therapeutic agents.

The present disclosure is also directed to methods of use or uses of the compounds of formula (I). In embodiments, compounds of formula (I) are used to treat or ameliorate hepatitis B virus (HBV) infection, to increase inhibition of HBV production, to inhibit HBV capsid assembly or other HBV viral replication processes, or their production. Used to block things. The method comprises administering to a subject in need of such method an effective amount of at least one compound of formula (I), a pharmaceutically acceptable salt of a compound of formula (I), Including administering pharmaceutically acceptable prodrugs and pharmaceutically active metabolites of compounds of formula (I). The detailed description describes additional embodiments of treatment methods.

It is an object of the present disclosure to overcome or ameliorate at least one of the prior art and/or shortcomings of the prior art, or to provide a useful alternative thereto. Further embodiments, features and advantages of the present disclosure will become apparent from the detailed description below and through practice of the disclosed subject matter.

Further embodiments, features and advantages of the disclosed subject matter will become apparent from the following detailed description of such disclosure and through practice thereof. For brevity, the publications, including patents, referred to in this specification are hereby incorporated by reference.

As used herein, compounds of formula (I), including compounds of formula (IA) and formula (II), their pharmaceutically acceptable salts, pharmaceutically acceptable prodrugs and compounds disclosed herein A pharmaceutically active metabolite of the compound is provided.

（式中、
Ｒ^１は、Ｆ、ＯＨ及びＣ_１～６アルキルからなる群から選択され、ここで、アルキルは、ＯＨで任意選択的に置換されており；
Ｒ^２は、Ｂｒ、ＣＮ及びＣ_１～４ハロアルキルからなる群から選択され；
Ｒ^３は、Ｈ又はＦであり；
Ｒ^４は、Ｈ又はＣ_１～４アルキルであり；
Ｘは、Ｏ、Ｓ、Ｓ＝Ｏ及びＳＯ_２からなる群から選択され；及び
Ｙは、ＣＨ、ＣＦ及びＮからなる群から選択される）
の化合物及びその薬学的に許容される塩、立体異性体、同位体変異体、Ｎ－オキシド又は溶媒和物が本明細書において提供される。 In one aspect, formula (I)

(In the formula,
R ¹ is selected from the group consisting of F, OH and C _1-6 alkyl, wherein alkyl is optionally substituted with OH;
R ² is selected from the group consisting of Br, CN and C _1-4 haloalkyl;
R ³ is H or F;
R ⁴ is H or C _1-4 alkyl;
X is selected from the group consisting of O, S, S ₌ O and SO2; and Y is selected from the group consisting of CH, CF and N)
and pharmaceutically acceptable salts, stereoisomers, isotopic variants, N-oxides or solvates thereof are provided herein.

In embodiments, the compound of formula (I) is
R ¹ is selected from the group consisting of F, OH and C _1-6 alkyl;
R ² is selected from the group consisting of Br, CN and C _1-4 haloalkyl;
R ³ is H or F; R ⁴ is H or C _1-4 alkyl;
X is selected from the group consisting of O, S, S ₌ O and SO2; and Y is a compound selected from the group consisting of CH, CF and N.

In embodiments, the compounds of formula (I) are those wherein R ¹ is OH.

In embodiments, the compounds of formula (I) are those in which R ¹ is F.

In embodiments, the compounds of formula (I) are those wherein R ¹ is C _1-6 alkyl.

In an embodiment, the compounds of formula (I) are those wherein R ¹ is hydroxymethyl.

In an embodiment, the compounds of formula (I) are those wherein ^R2 is Br, CN or _CF3 .

In embodiments, the compounds of formula (I) ^are those in which R3 is H.

In embodiments, the compounds of formula (I) ^are those in which R3 is F.

In an embodiment, the compounds of formula (I) are those in which ^R4 is H.

In an embodiment, the compounds of formula (I) are those wherein ^R4 is _CH3 .

In embodiments, the compounds of formula (I) are those in which Y is N.

In an embodiment, the compounds of formula (I) are those in which Y is CF.

In embodiments, the compounds of formula (I) are those in which Y is CH.

In embodiments, the compounds of formula (I) are those in which X is O.

In embodiments, the compounds of formula (I) are those in which X is S.

In embodiments, the compounds of formula (I) are those in which X is S=O.

In an embodiment, the compounds of _formula (I) are those in which X is SO2.

が３－シアノ－４－フルオロフェニル、４－フルオロ－３－（トリフルオロメチル）フェニル、３－シアノ－２，４－ジフルオロフェニル、３－ブロモ－２，４－ジフルオロフェニル、２－（ジフルオロメチル）－３－フルオロピリジン－４－イル又は２－ブロモ－３－フルオロピリジン－４－イルである化合物である。 In embodiments, the compound of formula (I) is

is 3-cyano-4-fluorophenyl, 4-fluoro-3-(trifluoromethyl)phenyl, 3-cyano-2,4-difluorophenyl, 3-bromo-2,4-difluorophenyl, 2-(difluoromethyl )-3-fluoropyridin-4-yl or 2-bromo-3-fluoropyridin-4-yl.

が３－シアノ－４－フルオロフェニルである化合物である。 In embodiments, the compound of formula (I) is

is 3-cyano-4-fluorophenyl.

並びにそれらの薬学的に許容される塩、Ｎ－オキシド又は溶媒和物からなる群から選択される化合物である。 Yet another embodiment of the present disclosure provides:

and pharmaceutically acceptable salts, N-oxides or solvates thereof.

（式中、
Ｒ^１は、Ｆ、ＯＨ及びＣ_１～６アルキルからなる群から選択され、ここで、アルキルは、ＯＨで任意選択的に置換されており；
Ｒ^２は、Ｂｒ、ＣＮ及びＣ_１～４ハロアルキルからなる群から選択され；
Ｒ^３は、Ｈ又はＦであり；
Ｒ^４は、Ｈ又はＣ_１～４アルキルであり；
Ｘは、Ｏ、Ｓ、Ｓ＝Ｏ及びＳＯ_２からなる群から選択され；及び
Ｙは、ＣＨ、ＣＦ及びＮからなる群から選択される）
の少なくとも１つの化合物及び式（Ｉ）の化合物の薬学的に許容される塩、立体異性体、同位体変異体、Ｎ－オキシド又は溶媒和物と；
（Ｂ）少なくとも１つの薬学的に許容される賦形剤と
を含む医薬組成物も開示される。 Pharmaceutical Compositions As used herein,
(A) Formula (I):

(In the formula,
R ¹ is selected from the group consisting of F, OH and C _1-6 alkyl, wherein alkyl is optionally substituted with OH;
R ² is selected from the group consisting of Br, CN and C _1-4 haloalkyl;
R ³ is H or F;
R ⁴ is H or C _1-4 alkyl;
X is selected from the group consisting of O, S, S ₌ O and SO2; and Y is selected from the group consisting of CH, CF and N)
and a pharmaceutically acceptable salt, stereoisomer, isotopic variant, N-oxide or solvate of a compound of formula (I);
Also disclosed are pharmaceutical compositions comprising (B) at least one pharmaceutically acceptable excipient.

並びにそのような化合物の任意の薬学的に許容される塩、Ｎ－オキシド若しくは溶媒和物又はそのような化合物の任意の薬学的に許容されるプロドラッグ若しくはそのような化合物の任意の薬学的に活性な代謝産物からなる群から選択される少なくとも１つの化合物とを含む医薬組成物である。 One embodiment of the present disclosure provides at least one pharmaceutically acceptable excipient,

and any pharmaceutically acceptable salts, N-oxides or solvates of such compounds or any pharmaceutically acceptable prodrugs of such compounds or any pharmaceutically acceptable compounds of such compounds. and at least one compound selected from the group consisting of active metabolites.

In embodiments, the pharmaceutical composition comprises at least one additional active or therapeutic agent. Additional active therapeutic agents include, for example, anti-HBV agents such as HBV polymerase inhibitors, interferons, viral entry inhibitors, viral maturation inhibitors, capsid assembly regulators, reverse transcriptase inhibitors, immunomodulatory agents such as TLR agonists. or any other agent that affects the HBV life cycle and/or outcome of HBV infection. Active agents of the disclosure are used alone or in combination with one or more additional active agents to prepare pharmaceutical compositions of the disclosure.

As used herein, the term "composition" or "pharmaceutical composition" refers to a mixture of at least one compound useful within the present disclosure and a pharmaceutically acceptable carrier. A pharmaceutical composition facilitates administration of the compound to a patient or subject. There are many techniques of administering compounds in the art, including but not limited to intravenous, oral, aerosol, parenteral, intraocular, intrapulmonary and topical administration.

As used herein, the term "pharmaceutically acceptable carrier" refers to a compound useful within the scope of this disclosure, within or within a patient, so as to be capable of performing its intended function. pharmaceutically acceptable liquid or solid fillers, stabilizers, dispersants, suspending agents, diluents, excipients, thickeners, solvents or encapsulating materials involved in carrying or transporting It means material, composition or carrier. Typically, such constructs are carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of formulations containing compounds useful within the present disclosure and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer; and other non-toxic, compatible substances used in pharmaceutical formulations.

As used herein, a "pharmaceutically acceptable carrier" is any coating that is compatible with the activity of the compounds useful within the scope of this disclosure and is physiologically acceptable to the patient. Also included are agents, antibacterial and antifungal agents, absorption delaying agents, and the like. Supplementary active compounds can also be incorporated into the compositions. A "pharmaceutically acceptable carrier" can further include pharmaceutically acceptable salts of the compounds useful within this disclosure. Other additional ingredients that can be included in pharmaceutical compositions used in the practice of the present disclosure are known in the art, see, for example, Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton , PA). This document is incorporated herein by reference.

A "pharmaceutically acceptable excipient" is a substance that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject, e.g., a pharmacological composition means an inert substance compatible with, or used as a vehicle, carrier, or diluent to facilitate administration of a drug. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starches, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

Pharmaceutical composition delivery forms containing one or more dosage units of the active agent can be prepared using suitable pharmaceutical excipients and using compounding techniques known or available to those of ordinary skill in the art. . In the methods of the present invention, compositions may be administered by any suitable delivery route, such as oral, parenteral, rectal, topical or ocular routes or by inhalation.

Formulations can be in the form of tablets, capsules, sachets, dragees, powders, granules, lozenges, powders for reconstitution, liquid formulations or suppositories. Preferably, the compositions are formulated for intravenous infusion, topical administration or oral administration.

For oral administration, the compounds of the disclosure can be provided in tablet or capsule form, or as a solution, emulsion or suspension. To prepare an oral composition, the compound may be added, for example, in doses of about 0.05 to about 100 mg/kg daily, about 0.05 to about 35 mg/kg daily, or about 0.1 to about 10 mg/kg daily. It can be prepared to give a dose. For example, a total daily dosage of about 5 mg to 5 g per day can be achieved by administration once, twice, three times or four times daily.

For oral tablets, the present disclosure mixed with pharmaceutically acceptable excipients such as inert diluents, tablet disintegrating agents, binders, lubricants, sweeteners, flavoring agents, coloring agents and preservatives. may include compounds according to Suitable inert fillers include sodium and calcium carbonates, sodium and calcium phosphates, lactose, starches, sugars, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol, and the like. Exemplary liquid oral excipients include ethanol, glycerol, water and the like. Starch, polyvinyl-pyrrolidone (PVP), sodium starch glycolate, microcrystalline cellulose and alginic acid are suitable tablet disintegrants. Binding agents may include starch and gelatin. The lubricant, if present, can be magnesium stearate, stearic acid or talc. If desired, the tablets can be coated with a substance such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract, or can be coated with an enteric coating.

Capsules for oral administration include hard and soft gelatin capsules. To prepare hard gelatin capsules, the compounds of the disclosure can be mixed with a solid, semi-solid, or liquid diluent. Soft gelatin capsules can be prepared by mixing a compound of the present disclosure with water, an oil such as peanut oil or olive oil, liquid paraffin, mixtures of mono- and diglycerides of short chain fatty acids, polyethylene glycol 400 or propylene glycol.

Liquids for oral administration may be in the form of suspensions, solutions, emulsions or syrups, or may be lyophilized or lyophilized for reconstitution with water or other suitable vehicle before use. It can be presented as a dry formulation. Such liquid compositions optionally contain pharmaceutically acceptable excipients such as suspending agents (e.g., sorbitol, methylcellulose, sodium alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel, etc.). agents; non-aqueous vehicles such as oils (eg almond or coconut oil), propylene glycol, ethyl alcohol or water; preservatives (eg methyl p-hydroxybenzoate or propyl p-hydroxybenzoate or sorbic acid); lecithin. and, optionally, flavoring or coloring agents.

The active agents of this disclosure can also be administered by parenteral routes. For example, the compositions may be prepared as suppositories for rectal administration. For parenteral use, including intravenous, intramuscular, intraperitoneal or subcutaneous routes, the compounds of this disclosure are in sterile aqueous solutions or suspensions, buffered to appropriate pH and isotonicity, or parenterally acceptable. can be provided in an oil Suitable aqueous vehicles include Ringer's solution and isotonic sodium chloride. Such forms include unit dosage forms such as ampoules or disposable injection devices, multi-dose forms such as vials from which appropriate doses can be extracted, or solid forms that can be used to prepare injectable formulations. Or it can be presented in a pre-concentrate. A specific injection dose can range from about 1-1000 μg/kg/minute of compound mixed with a pharmaceutical carrier over a period ranging from minutes to days.

For topical administration, the compounds may be mixed with a pharmaceutical carrier at a concentration of about 0.1% to about 10% drug to vehicle. Another mode of administration of the compounds of the present disclosure may utilize patch formulations to affect transdermal delivery.

Alternatively, the compounds of this disclosure can be administered in the methods of this disclosure by inhalation, by nasal or oral routes, eg, in a spray formulation additionally containing a suitable carrier.

Methods of Use The compounds disclosed herein are useful for treating and preventing HBV infection in subjects, including human subjects.

In a non-limiting aspect, these compounds are capable of (i) modulating or disrupting HBV assembly and other HBV core protein functions required for HBV replication or generation of infectious particles, or (ii) infectious viral particles. or (iii) interact with the HBV capsid and act on defective viral particles with weak infectivity or replication capacity acting as capsid assembly regulators. Specifically, without being bound by any particular mechanism of action, the compounds of the present disclosure disrupt and promote normal viral capsid assembly/disassembly of immature or mature particles in HBV therapy. , reduce, slow and/or inhibit, thereby inducing abnormal capsid morphology leading to antiviral effects such as virion assembly and/or disassembly, virion maturation, viral release and/or infection of target cells. considered useful by Compounds of the present disclosure may function as disruptors of capsid assembly interacting with mature or immature viral capsids, disrupting capsid stability and thus affecting its assembly and/or disassembly. The compounds of the present disclosure may disrupt protein folding and/or salt bridges necessary for viral capsid function and/or normal morphology, thereby disrupting and/or promoting capsid assembly and/or disassembly. Compounds of the present disclosure bind to the capsid and alter the metabolism of cellular polyproteins and precursors, thereby inducing cytotoxicity and death of infected cells. can lead to significant accumulation. Compounds of the present disclosure may cause hypoformation of capsids with optimal stability and affect efficient uncoating and/or degradation of viruses (eg, during infectivity). Compounds of the present disclosure may disrupt and/or promote capsid assembly and/or disassembly when the capsid proteins are immature. Compounds of the present disclosure may disrupt and/or promote capsid assembly and/or disassembly when capsid proteins are mature. Compounds of the present disclosure may disrupt and/or promote capsid assembly and/or disassembly during viral infection, which may further attenuate HBV viral infectivity and/or reduce viral load. Disruption, promotion, inhibition, retardation and/or reduction of capsid assembly and/or disassembly by compounds of the present disclosure can eradicate the virus from the host organism. Eradication of HBV from a subject by the compounds of the present disclosure advantageously obviates the need for and/or shortens the duration of long-term long-term therapy.

An additional embodiment of the present disclosure is a method of treating a subject suffering from HBV infection, comprising administering to the subject in need of such treatment an effective amount of at least one compound of Formula (I) It is a method that includes

In another aspect, provided herein is a method of reducing viral load associated with HBV infection in an individual in need thereof, comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable A method is provided comprising administering to the individual a salt of a given salt.

In another aspect, provided herein is a method of reducing recurrence of HBV infection in an individual in need thereof, comprising: a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof; is provided to the individual.

In another aspect, provided herein is a method of inhibiting or reducing the formation or presence of HBV DNA-containing particles or HBV RNA-containing particles in an individual in need thereof, comprising a therapeutically effective amount of formula (I) or a pharmaceutically acceptable salt thereof to the individual.

In another aspect, provided herein is a method of reducing the adverse physiological effects of HBV infection in an individual in need thereof, comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically A method is provided comprising administering an acceptable salt to the individual.

In another aspect, provided herein is a method of inducing remission of liver damage from HBV infection in an individual in need thereof, comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically A method is provided comprising administering an acceptable salt to the individual.

In another aspect, provided herein is a method of reducing the physiological effects of long-term antiviral therapy against HBV infection in an individual in need thereof, comprising a therapeutically effective amount of a compound of formula (I) or A method is provided comprising administering a pharmaceutically acceptable salt to the individual.

In another aspect, provided herein is a method of prophylactic treatment of HBV infection in an individual in need thereof, wherein the individual is suffering from subclinical HBV infection, the method comprising: A method is provided comprising administering to the individual a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof.

In embodiments, compounds of the present disclosure are suitable for monotherapy. In embodiments, compounds of the present disclosure are effective against natural or natural HBV strains. In embodiments, compounds of the present disclosure are effective against HBV strains that are resistant to currently known drugs.

In another embodiment, the compounds provided herein can be used in methods of modulating (eg, inhibiting or disrupting) HBV cccDNA activity, stability, function and viral replication properties.

In yet another embodiment, compounds of the present disclosure may be used in methods of reducing or preventing the formation of HBV cccDNA.

In another embodiment, the compounds provided herein can be used in methods of modulating (eg, inhibiting or disrupting) the activity of HBV cccDNA.

In yet another embodiment, compounds of the present disclosure can be used in methods of reducing the formation of HBV cccDNA.

In another embodiment, compounds of the present disclosure may be used in methods of modulating, inhibiting or disrupting the production or release of HBV RNA particles from within infected cells.

In further embodiments, the total amount (or concentration) of HBV RNA particles is modulated. In preferred embodiments, the total amount of HBV RNA is reduced.

In another embodiment, the methods provided herein are HBV polymerase inhibitors, interferons, viral entry inhibitors, viral maturation inhibitors, distinct capsid assembly regulators, antiviral agents of distinct or unknown mechanism. A greater or more rapid reduction in viral load in the individual compared to administration of a compound selected from the group consisting of the compound and any combination thereof.

In another embodiment, the methods provided herein use HBV polymerase inhibitors, interferons, viral entry inhibitors, viral maturation inhibitors, different capsid assembly regulators, antiviral compounds with different or unknown mechanisms. and a lower incidence of viral mutation and/or viral resistance compared to administration of a compound selected from the group consisting of and combinations thereof.

In another embodiment, the methods provided herein further comprise administering to the individual at least one HBV vaccine, nucleoside HBV inhibitor, interferon, or any combination thereof.

In one aspect, provided herein is a method of treating HBV infection in an individual in need thereof, comprising administering a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, to the individual. and reducing HBV viral load by further administering to the individual a therapeutically effective amount of an HBV vaccine.

An additional embodiment of the present disclosure is a method of treating a subject suffering from HBV infection, comprising administering to the subject in need of such treatment an effective amount of at least one compound of Formula (I) It is a method that includes

In another aspect, provided herein is a method of reducing viral load associated with HBV infection in an individual in need thereof, comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable A method is provided comprising administering to the individual a salt of a given salt.

In another aspect, provided herein is a method of reducing recurrence of HBV infection in an individual in need thereof, comprising: a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof; is provided to the individual.

In another aspect, provided herein is a method of inhibiting or reducing the formation or presence of HBV DNA-containing particles or HBV RNA-containing particles in an individual in need thereof, comprising a therapeutically effective amount of formula (I) or a pharmaceutically acceptable salt thereof to the individual.

In another aspect, provided herein is a method of reducing the adverse physiological effects of HBV infection in an individual in need thereof, comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically A method is provided comprising administering an acceptable salt to the individual.

In another aspect, provided herein is a method of inducing remission of liver damage from HBV infection in an individual in need thereof, comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically A method is provided comprising administering an acceptable salt to the individual.

In another aspect, provided herein is a method of reducing the physiological effects of long-term antiviral therapy against HBV infection in an individual in need thereof, comprising a therapeutically effective amount of a compound of formula (I) or A method is provided comprising administering a pharmaceutically acceptable salt to the individual.

In another aspect, provided herein is a method of prophylactic treatment of HBV infection in an individual in need thereof, wherein the individual is suffering from subclinical HBV infection, the method comprising: A method is provided comprising administering to the individual a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof.

In one embodiment, the methods provided herein further comprise monitoring the subject's HBV viral load, wherein the method is performed over a period of time such that the HBV virus is undetectable.

Combinations Provided herein are combinations of one or more compounds of the present disclosure and at least one therapeutic agent. In embodiments, the methods provided herein can further comprise administering at least one additional therapeutic agent to the individual. In embodiments, compounds of the present disclosure are suitable for use in combination therapy. Compounds of the present disclosure may be useful in combination with one or more additional compounds useful in treating HBV infection. These additional compounds may include compounds of the disclosure and/or compounds known to treat, prevent or alleviate the symptoms or effects of HBV infection.

In an exemplary embodiment, the additional active ingredient is another HBV capsid assembly regulator or a compound that is active against a specific condition or disorder associated with HBV infection or another target associated with HBV infection itself. Active ingredients known or found to be effective in the treatment of conditions or disorders associated with HBV infection. A combination may increase efficacy (e.g., by including in combination a compound that enhances the potency or effectiveness of an active agent according to the present disclosure), reduce one or more side effects, or reduce an active agent according to the present disclosure. can help reduce the need for In further embodiments, the methods provided herein include at least one additional treatment required to achieve similar results as in prophylactically treating HBV infection in an individual in need thereof. Allows administration of at least one additional therapeutic agent at a lower dose or frequency compared to administering the drug alone.

Such compounds include HBV combinations, HBV vaccines, HBV DNA polymerase inhibitors, immunomodulatory agents, toll-like receptor (TLR) modulators, interferon alpha receptor ligands, hyaluronidase inhibitors, hepatitis B surface antigen ( HBsAg) inhibitors, cytotoxic T lymphocyte-associated protein 4 (ipi4) inhibitors, cyclophilin inhibitors, HBV virus entry inhibitors, antisense oligonucleotides targeting viral mRNAs, short interfering RNA (siRNA) and ddRNAi Endonuclease modulators, ribonucleotide reductase inhibitors, HBV E antigen inhibitors, covalent closed circular DNA (cccDNA) inhibitors, farnesoid X receptor agonists, HBV antibodies, CCR2 chemokine antagonists, thymosin agonists, cytokines, nuclear proteins Modulators, retinoic acid-inducible gene 1 stimulators, NOD2 stimulators, phosphatidylinositol 3-kinase (PI3K) inhibitors, indoleamine-2,3-dioxygenase (IDO) pathway inhibitors, PD-1 inhibitors, PD - L1 inhibitors, recombinant thymosin alpha-1, Bruton's Tyrosine Kinase (BTK) inhibitors, KDM inhibitors, HBV replication inhibitors, Arginase inhibitors and affecting the HBV life cycle and/or consequences of HBV infection Any other drug or combination thereof that affects

In embodiments, the compounds of the disclosure are HBV polymerase inhibitors, immunomodulators, interferons such as peginterferon, viral entry inhibitors, viral maturation inhibitors, capsid assembly regulators, reverse transcriptase inhibitors, cyclophilin/TNF inhibitors immunomodulators such as TLR agonists, HBV vaccines and any other drug that affects the HBV life cycle and/or the outcome of HBV infection, or combinations thereof.

In particular, compounds of the present disclosure may be used in combination with one or more agents (or salts thereof) selected from the group consisting of:
HBV reverse transcription inhibitors and DNA and RNA polymerase inhibitors such as, but not limited to, lamivudine (3TC, Zeffix, Heptovir, Epivir and Epivir-HBV), entecavir (Baraclude, Entavir), adefovir dipivoxil (Hepsara, Preveon , bis-POM PMEA), tenofovir disoproxil fumarate (Viread, TDF or PMPA);
interferons such as, but not limited to, interferon alpha (IFN-α), interferon beta (IFN-β), interferon lambda (IFN-λ) and interferon gamma (IFN-γ);
viral entry inhibitors;
viral maturation inhibitors;
capsid assembly regulators described in literature such as, but not limited to, BAY 41-4109;
reverse transcriptase inhibitors;
immunomodulatory agents such as TLR agonists; and AT-61 ((E)-N-(1-chloro-3-oxo-1-phenyl-3-(piperidin-1-yl)propa-1), including but not limited to -en-2-yl)benzamide), AT-130 ((E)-N-(1-bromo-1-(2-methoxyphenyl)-3-oxo-3-(piperidin-1-yl)prop-1 -en-2-yl)-4-nitrobenzamide) and similar analogues with different or unknown mechanisms.

In embodiments, the additional therapeutic agent is interferon. The term "interferon" or "IFN" refers to any member of a family of highly homologous species-specific proteins that inhibit viral replication and cell proliferation and modulate immune responses. Human interferons are divided into three classes: type I, which includes interferon alpha (IFN-α), interferon beta (IFN-β), and interferon omega (IFN-ω); type II, which includes interferon gamma (IFN-γ); and interferon lambda. It is classified as type III, including (IFN-λ). Recombinant interferons that have been developed and are commercially available are encompassed by the term "interferon" as used herein. Subtypes of interferon, such as chemically modified or mutated interferons, are also encompassed by the term "interferon" as used herein. Chemically modified interferons include pegylated interferons and glycosylated interferons. Examples of interferons also include interferon alpha-2a, interferon alpha-2b, interferon alpha-n1, interferon beta-1a, interferon beta-1b, interferon lambda-1, interferon lambda-2 and interferon lambda-3, It is not limited to these. Examples of pegylated interferons include pegylated interferon alpha-2a and pegylated interferon alpha-2b.

Thus, in one embodiment, the compound of Formula I is selected from the group consisting of interferon alpha (IFN-α), interferon beta (IFN-β), interferon lambda (IFN-λ) and interferon gamma (IFN-γ). It can be administered in combination with interferon. In one particular embodiment, the interferon is interferon alpha-2a, interferon alpha-2b or interferon alpha-n1. In another specific embodiment, interferon alpha-2a or interferon alpha-2b is pegylated. In preferred embodiments, the interferon alpha-2a is peginterferon alpha-2a (PEGASYS).

In another embodiment, the additional therapeutic agent is selected from immunomodulatory or immunostimulatory therapy, including biological agents belonging to the interferon class.

Additionally, additional therapeutic agents may be agents that disrupt the function of other essential viral proteins or host proteins required for HBV replication or persistence.

In another embodiment, the additional therapeutic agent is an antiviral agent that blocks viral entry or maturation or targets HBV polymerase, such as nucleoside or nucleotide or non-nucleoside (t)polymerase inhibitors. be. In further embodiments of combination therapy, the reverse transcriptase inhibitor and/or DNA and/or RNA polymerase inhibitor is zidovudine, didanosine, zalcitabine, ddA, stavudine, lamivudine, abacavir, emtricitabine, entecavir, aplicitabine, atevirapine, ribavirin, acyclovir, famciclovir, valacyclovir, ganciclovir, valganciclovir, tenofovir, adefovir, PMPA, cidofovir, efavirenz, nevirapine, delavirdine or etravirine.

In one embodiment, the additional therapeutic agent is an immunomodulatory agent that induces an innate limited immune response that results in induction of an immune response against a distinct virus. In other words, the immunomodulatory agent regulates antigen-presenting cell maturation, T-cell proliferation and cytokine release (e.g. IL-12, IL-18, IFN-alpha, IFN-beta and IFN-gamma and TNF-alpha among others). Bring.

In further embodiments, the additional therapeutic agent is a TLR modifying agent or TLR agonist such as a TLR-7 agonist or a TLR-9 agonist. In a further embodiment of combination therapy, the TLR-7 agonists are SM360320 (9-benzyl-8-hydroxy-2-(2-methoxy-ethoxy)adenine) and AZD8848 ([3-({[3-(6-amino -2-butoxy-8-oxo-7,8-dihydro-9H-purin-9-yl)propyl][3-(4-morpholinyl)propyl]amino}methyl)phenyl]methyl acetate); be.

In any of the methods provided herein, the method further comprises administering to the individual at least one HBV vaccine, nucleoside HBV inhibitor, interferon, or any combination thereof. In one embodiment, the HBV vaccine is at least one of RECOMBIVAX HB, ENGERIX-B, ELOVAC B, GENEVAC-B or SHANVAC B.

In another aspect, provided herein is a method of treating HBV infection in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of the present disclosure, alone or in combination with reverse transcriptase. and reducing HBV viral load by further administering to the individual a therapeutically effective amount of an HBV vaccine. Reverse transcriptase inhibitors include zidovudine, didanosine, zalcitabine, ddA, stavudine, lamivudine, abacavir, emtricitabine, entecavir, apricitabine, atevirapine, ribavirin, acyclovir, famciclovir, valacyclovir, ganciclovir, valganciclovir, tenofovir, adefovir, PMPA, It may be one of cidofovir, efavirenz, nevirapine, delavirdine or etravirine.

For any combination therapy described herein, synergistic effects can be measured by, for example, the sigmoidal E _max equation (Holford & Scheiner, 19981, Clin. Pharmacokinet. 6:429-453), the Loewe additivity equation (Loewe & Muischnek, 1926, Arch. Exp. Pathol Pharmacol. 114:313-326) and the median effect equation (Chou & Talalay, 1984, Adv. Enzyme Regul. 22:27-55). Each of the equations referred to above can be applied to the experimental data to produce a corresponding graph to aid in assessing the effects of drug combinations. The corresponding graphs associated with the equations referred to above are concentration-effect curves, isobologram curves and combination exponential curves, respectively.

Definitions Listed below are definitions of various terms used to describe this disclosure. These definitions apply to the terms used throughout the specification and claims unless otherwise limited individually or as part of a larger group in specific instances.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the applicable art. In general, the nomenclature and laboratory procedures in cell culture, molecular genetics, organic chemistry and peptide chemistry used herein are those well known and commonly used in the art.

As used herein, the articles "a" and "an" refer to one or more (ie, at least one) of the grammatical objects of the article. By way of example, "an element" means one element or more than one element. Furthermore, the use of the term "includes" and other forms such as "includes", "includes" and "included" is not limiting.

As used in the specification and claims, the term "comprising" can include "consisting of" and "consisting essentially of" embodiments. The terms “comprising,” “includes,” “has,” “has,” “can,” “contains,” and variations thereof, as used herein, refer to the specified component/ It shall be an open-ended transitional phrase, term or word that requires the presence of a step and allows the presence of other components/steps. However, such statements describe the composition or process as "consisting of" and "consisting essentially of" the recited compounds, i.e., together with any pharmaceutically acceptable carrier, the specified It should also be interpreted as allowing only the compound to be present and excluding other compounds. All ranges disclosed herein are inclusive of the recited endpoints and are individually combinable (eg, a range “50 mg to 300 mg” includes the endpoints 50 mg and 300 mg and all intermediate values). The endpoints of the ranges and any values disclosed herein are not limited to precise ranges or values, but are vague enough to include values approximating these ranges and/or values.

As used herein, the term expressing approximation may be applied to modify any quantitative expression that may change without changing the underlying function to which it is associated. Thus, values modified by terms such as "substantially" can sometimes not be limited to the precise value specified. In at least some examples, the approximating term may correspond to the precision of an instrument for measuring the value.

The term "alkyl" represents straight or branched chain alkyl groups having 1 to 12 carbon atoms in the chain. Examples of alkyl groups include methyl (Me, which can be structurally depicted by the symbol "/"), ethyl (Et), n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl ( tBu), pentyl, isopentyl, tert-pentyl, hexyl, isohexyl and groups that would be considered equivalent to any one of the above examples in light of the knowledge of those skilled in the art and the teachings provided herein. is mentioned. The term C _1-4 alkyl as used herein refers to a straight or branched chain alkyl group having 1 to 4 carbon atoms in the chain. The term C _1-6 alkyl as used herein refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms in the chain.

The term "heteroaryl" refers to a monocyclic or fused bicyclic heterocycle having 3 to 9 ring atoms per heterocycle (carbon atoms and up to 4 heterocycles selected from nitrogen, oxygen and sulfur). ring structure having ring atoms selected from atoms). Illustrative examples of heteroaryl groups include the following entities in the form of appropriately linked moieties.

Those skilled in the art will recognize that the species of heteroaryl groups listed or exemplified above are not exclusive and that species within these defined terms may additionally be selected.

The term "cyano" refers to the -CN group.

The term "halo" represents chloro, fluoro, bromo or iodo.

The term "perhaloalkyl" or "haloalkyl" refers to a straight or branched chain alkyl group having 1-6 carbon atoms in the chain and optionally replacing the hydrogens with halogens. The term "C _1-4 haloalkyl" as used herein refers to a straight or branched chain alkyl group having 1 to 4 carbon atoms in the chain and optionally replacing hydrogen with halogen. point to The term "C _1-6 haloalkyl" as used herein refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms in the chain and optionally replacing hydrogen with halogen. point to Examples of "perhaloalkyl" or "haloalkyl" groups include trifluoromethyl _{( CF3} ), difluoromethyl ( _CF2H ), monofluoromethyl ( _CH2F ), pentafluoroethyl ( _CF2CF3 ), tetrafluoroethyl (CHFCF ₃ ), monofluoroethyl (CH ₂ CH ₂ F), trifluoroethyl (CH ₂ CF ₃ ), tetrafluorotrifluoromethylethyl (-CF(CF ₃ ) ₂ ) and those skilled in the art and Included are groups that are considered equivalent to any of the foregoing examples in light of the teachings provided herein.

The term "substituted" means that the specified group or moiety has one or more substituents. The term "unsubstituted" means that the specified group has no substituents. The term "optionally substituted" means that the specified group is unsubstituted or substituted with one or more substituents. When the term "substituted" is used to describe a structural system, substitution shall occur at any position on the system where valences permit. Unless it is expressly stated that a particular moiety or group is optionally substituted or substituted with any particular substituent, such moiety or group is meant to be unsubstituted. It should be understood that this is intended.

The terms "para", "meta" and "ortho" have their art-understood meanings. Thus, for example, a fully substituted phenyl group has both "ortho" (o) positions adjacent to the point of attachment of the phenyl ring, both "meta" (m) positions and one "para" position opposite the point of attachment. ” (p) has a substituent at the position. To make the position of the substituents on the phenyl ring more clear, two different ortho positions are designated as ortho and ortho' and two different meta positions are designated as meta and meta' positions, as exemplified below. do.

When referring to substituents on a pyridyl group, the terms "para", "meta" and "ortho" refer to the position of the substituent relative to the point of attachment of the pyridyl ring. For example, the structure below is illustrated as a 3-pyridyl with the X ¹ substituent at the ortho position, the X ² substituent at the meta position and the X ³ substituent at the para position.

To provide a more concise description, some of the quantitative expressions provided herein are not qualified with the term "about." Any quantity provided herein is intended to represent the actual given value, and whether or not the term "about" is explicitly used, and such given value refers to approximations to such given values that would reasonably be inferred based on the knowledge of one skilled in the art, including equivalents and approximations resulting from experimental and/or measurement conditions for It should be understood that it is also intended that When yields are given as percentages, such yields refer to the mass of the entity for which the yield is given relative to the maximum amount of the same entity that can be obtained under specified stoichiometric conditions. Concentrations given in percentages refer to mass ratios, unless otherwise indicated.

The terms "buffered" or "buffered" fluids are used interchangeably herein according to their standard meanings. Buffers are used to control the pH of media and their selection, use and function are known to those skilled in the art. For example, G. et al., which describes, inter alia, how the concentrations of buffers and buffer components relate to the pH of the buffer. D. Considine, ed. , Van Nostrand's Encyclopedia of Chemistry, p. ²⁶¹ , 5th ed. (2005). For example, a buffer is obtained by adding MgSO4 and _NaHCO3 to the solution in a ratio of ₁₀ :1 (w/w) to maintain the pH of the solution at about 7.5.

Any formula given herein is intended to represent a compound having a structure depicted by the structural formula and specific variations or forms. In particular, compounds of any formula given herein may have asymmetric centers and therefore exist in different enantiomeric forms. All optical isomers of compounds of general formula and mixtures thereof are considered to be included within the scope of the formula. Thus, any formula given herein is intended to represent the racemate, one or more enantiomeric forms, one or more diastereomeric forms, one or more atropisomeric forms and mixtures thereof. ing. Additionally, certain structures may exist as geometric isomers (ie cis and trans isomers), tautomers or atropisomers.

It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed "isomers."

Stereoisomers that are not mirror images of one another are termed "diastereomers" and stereoisomers that are non-superimposable mirror images of one another are termed "enantiomers". When a compound has an asymmetric center, for example, it can be attached to four different groups, and enantiomeric pairs are possible. Enantiomers can be characterized by the absolute configuration of their chiral centers, by the Kahn and Prelogue R and S precedence rules, or by the fact that the molecule is dextrorotatory or levorotatory (ie, (+)- or (-)-, respectively). isomers) are explained by the method of rotating the plane of polarized light. A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a "racemic mixture".

"Tautomers" refer to compounds that are structurally interchangeable forms of a particular compound and differ in rearrangements of hydrogen atoms and electrons. Therefore, the two structures can be in equilibrium due to the motion of pi-electrons and one atom (usually H). For example, enols and ketones are tautomers because they are rapidly interconverted by treatment with acid or base. Another example of tautomerism is the acyl and nitro forms of phenylnitromethane, which are similarly formed by treatment with acid or base.

Tautomeric forms may be important for a target compound to achieve optimum chemical reactivity and biological activity.

Compounds of the present disclosure may possess one or more asymmetric centers, and thus such compounds can be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof. .

Unless otherwise indicated, the description or naming of a particular compound within the specification and claims includes both individual enantiomers and mixtures thereof, whether racemic or not. is intended. Methods for determining stereochemistry and for separating stereoisomers are well known in the art.

Specific examples include chemical structures that are depicted as absolute enantiomers, but are intended to represent enantiopure substances whose configuration is not yet known. In these cases (R*) or (S*) are used in the designation to indicate that the absolute stereochemistry of the corresponding stereocenter is not yet known. Thus, a compound labeled (R*) refers to an enantiopure compound having the (R) or (S) absolute configuration. These structures are named using (R) and (S) when the absolute stereochemistry is confirmed.

は、本明細書に示した化学構造において同じ空間的配置を意味するものとして使用される。同様に、記号

は、本明細書に示した化学構造において同じ空間的配置を意味するものとして使用される。 symbol

are used to refer to the same spatial arrangement in the chemical structures shown herein. Similarly, the symbol

are used to refer to the same spatial arrangement in the chemical structures shown herein.

Additionally, any formula given herein refers to hydrates, solvates and polymorphs of such compounds and mixtures thereof, even if such forms are not explicitly recited. is intended. Certain compounds of formula (I) or pharmaceutically acceptable salts of compounds of formula (I) may be obtained as solvates. Solvates include those formed from the interaction or complexation of the compounds of the disclosure with one or more solvents, either in solution or in solid or crystalline form. In some embodiments the solvent is water and the solvate is a hydrate. Additionally, certain crystalline forms of a compound of formula (I) or a pharmaceutically acceptable salt of a compound of formula (I) may be obtained as co-crystals. In certain embodiments of the present disclosure, compounds of formula (I) were obtained in crystalline form. In other embodiments, the crystalline form of the compound of formula (I) was cubic in nature. In other embodiments, pharmaceutically acceptable salts of compounds of formula (I) were obtained in crystalline form. In still other embodiments, the compound of formula (I) was obtained in one of several polymorphic forms, as a mixture of crystalline forms, as polymorphic forms, or as an amorphous form. In other embodiments, the compounds of formula (I) convert between one or more crystalline and/or polymorphic forms in solution.

References to a compound herein refer to any one of the following: (a) the actually named form of such compound; and (b) the compound considered at the time the compound is named. Any of the forms of such compounds in the medium. For example, references herein to compounds such as R-COOH refer, for example, to any one of R-COOH _(s) , R-COOH _(sol) and R ^- COO- _(sol) . contain. In this example, R-COOH _(s) refers to the solid compound as it may be in, for example, a tablet or some other solid pharmaceutical composition or formulation; R-COOH _(sol) is the compound in solvent R—COO ⁻ _(sol) refers to whether such free form is derived from R—COOH, is derived from a salt thereof, or is considered to be the free form of a compound in an aqueous environment. It refers to the dissociated form of a compound in a solvent, irrespective of whether it is derived from any other entity that produces R ^- COO- after dissociation in the medium containing the solvent. In another example, a phrase such as "exposing an entity to a compound of formula R-COOH" refers to one or more forms of one or more compounds R-COOH present in a medium in which such exposure occurs. refers to the exposure of such entities in In yet another example, a phrase such as "reacting an entity with a compound of formula R--COOH" means that (a) one or more chemical entities of such entity are present in the medium in which such reaction occurs. (b) reacting one or more chemically significant forms of the compound R—COOH present in the medium in which such reaction occurs. In this regard, when such entities are in, for example, an aqueous environment, the compound R—COOH is present in such same medium, and thus the entities are represented by R—COOH _(aq) and/or R— It has been exposed to species such as COO ⁻ _(aq) . Here, the subscript "(aq)" stands for "aqueous" according to its conventional meaning in chemistry and biochemistry. A carboxylic acid functional group is chosen in these nomenclature examples, but this choice is not intended to be limiting, but merely illustrative. Analogous examples include other functional groups such as, but not limited to, hydroxyl, basic nitrogen members such as those in amines and any functional group that interacts or changes according to known methods in a medium containing the compound. It is understood that other groups may be provided. Such interactions and changes include, but are not limited to, dissociation, association, tautomerism, solvolysis, including hydrolysis, solvation including hydration, protonation and deprotonation. These interactions and changes in a given medium are known to those of ordinary skill in the art, so no further examples are provided in this regard here.

In another example, a zwitterionic compound is included herein by referring to a compound that is known to form a zwitterion, even if it is not explicitly named in zwitterionic form. be done. Terms such as zwitterions and their synonyms, zwitterionic compounds, are well-known and standard IUPAC-approved names that are part of a standard set of defined scientific names. In this regard, the name "zwitterion" is assigned the identification name CHEBI:27369 by the Chemical Entities of Biological Interest (ChEBI) Dictionary of Molecular Entities. As is generally well known, zwitterions or zwitterionic compounds are neutral compounds having formal unit charges of opposite signs. Sometimes these compounds are referred to by the term "inner salts". Other sources refer to these compounds as "dipole ions", although the latter term is considered a misnomer by still other sources. As a particular example, aminoethanoic acid (the amino acid glycine) has the formula H ₂ NCH ₂ COOH and in some media (in this case neutral media) in the form of zwitterion ⁺ H ₃ NCH ₂ COO ⁻ exist. Zwitterions, zwitterionic compounds, inner salts and dipoles in the known and well-established meanings of these terms will in each case be understood as such by those skilled in the art. are therefore included within the scope of this disclosure. Structures of zwitterionic compounds related to compounds of the present disclosure are not explicitly shown herein, as there is no need to name every embodiment that would be recognized by one of ordinary skill in the art. However, they are part of the embodiments of this disclosure. These interactions and changes in a given medium that result in various forms of a given compound are known to those skilled in the art, and therefore no further examples relating to this are provided herein.

Any formula given herein is also intended to represent unlabeled and isotopically labeled forms of the compound. Isotopically-labeled compounds have structures depicted by the formulas shown herein, except that one or more atoms are replaced by atoms having a selected atomic weight or mass number. Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine and iodine, such as ² H, ³ H, ¹¹ C, ¹³ C, respectively. ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ³⁶ Cl, ¹²⁵ I. Such isotopically labeled compounds may be used in metabolic studies (preferably using ¹⁴ C), kinetic studies (e.g. deuterium (i.e. D or ² H)), including drug or substrate tissue distribution assays, or tritium (i.e. T or ³ H))), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), or radiotherapy of patients. In particular, ¹⁸ F- or ¹¹ C-labeled compounds are preferred, especially in PET or SPECT studies. In addition, substitution with heavier isotopes, such as deuterium (i.e., ² H), may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or decreased dosage requirements. can bring Isotopically-labeled compounds of the present disclosure and their prodrugs can generally be prepared using readily available isotopically-labeled reagents in place of isotopically unlabeled reagents as disclosed in the schemes or examples and methods of preparation described below. can be prepared by carrying out the procedure described.

When referring to formulas presented herein, the selection of a particular moiety from a list of possible species for a particular variable is intended to provide for the same selection of species for the variable occurring elsewhere. not In other words, if a variable occurs more than once, the selection of species from a particular list is independent of the selection of species for the same variable elsewhere in the formula unless otherwise indicated.

In accordance with the above interpretative considerations regarding designations and nomenclature, explicit reference herein to a set is of chemical significance, and the practice of such a set unless otherwise indicated. A separate reference to a form and a reference to any possible embodiment of a subset of the explicitly mentioned set is meant.

As a first example of substituent terminology, if the S ¹ _example of a substituent is one of S ₁ and S ₂ and the S ² _example of a substituent is one of S ₃ and S ₄ , then these designations are , S ¹ _example is S ₁ and S ² _example is S ₃ ; S ¹ _example is S ₁ and S ² _example is S ₄ ; S ¹ _example is S ₂ and S ² _example is S ¹ _example is S ₂ and S ² _example is S ₄ ; and the equivalents of _each one of such choices. Thus, the short term "S ¹ _example is one of S ₁ and S ₂ and S ² _example is one of S ₃ and S ₄ " is used herein for brevity's sake. It is intended for use and is in no way limiting. The above first example of substituent terminology set forth in general terms is intended to illustrate the various substituent designations described herein. The above conventions provided herein for substituents apply where applicable to members such as R ¹ , R ² , R ³ , R ⁴ , PG, X and Y and any other general It also applies to generic substituent symbols.

Further, when more than one designation is given for any member or substituent, embodiments of the present disclosure contemplate various groupings that can be made from the recited designations and their equivalents taken independently. include. As a second example of substituent terminology, if a substituent S _example is described herein as being one of S ₁ , S ₂ and S ₃ , then the enumeration is such that S _example is S ₁ ; S _example is S ₃ ; S _example is one of S ₁ and S ₂ ; S _example is _one of S ₁ and S ₃ ; _{S example is} S ₂ and S ₃ ; S _example is one of S ₁ , S ₂ and S ₃ ; and S _example is any equivalent of each one of these choices. Thus, the short term "S _example is one of S ₁ , S ₂ and S ₃ " is used herein for brevity and is in no way limiting. . The above second example of substituent terminology being stated in general terms is intended to illustrate the various substituent designations described herein. The above conventions provided herein for substituents apply where applicable to members such as R ¹ , R ² , R ³ , R ⁴ , PG, X and Y and any other general It also applies to generic substituent symbols.

The term “C _i-j ” (where j>i), when applied herein to a class of substituents, means that any number of carbon members from i to j inclusive of i and j is It is intended to mean independently implemented embodiments of the present disclosure. By way of example, the terms C _1-4 refer to embodiments having 1 carbon member (C ₁ ), embodiments having 2 carbon members (C ₂ ), embodiments having 3 carbon members (C ₃ ) form and refers independently to embodiments with 4 carbon members (C ₄ ).

The term C _n-m alkyl means an aliphatic group, whether straight or branched, with a total of N carbon members in the chain such that n≦N≦m, where m>n. point to the chain. Any of the disubstituents referred to herein include the various bonding possibilities where two or more such possibilities are permitted. For example, reference to a disubstituent -AB-, where A≠B, is used herein to refer to A attached to the first substituent member and B attached to the second substituent member. It refers to such disubstituents with A attached to the second substituent member and also includes such disubstituents with B attached to the first substituent member.

The present disclosure also includes pharmaceutically acceptable salts of the compounds of formula (I), preferably the compounds described above and the specific compounds exemplified herein, and methods of treatment using such salts.

The term "pharmaceutically acceptable" has been or will be approved by regulatory agencies of federal or state governments or corresponding agencies in countries other than the United States for use in animals and more particularly in humans. , or those listed in the United States Pharmacopoeia or other generally accepted pharmacopoeia.

A "pharmaceutically acceptable salt" is a compound represented by Formula (I) that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject. is intended to mean a salt of the free acid or free base of A "pharmaceutically acceptable salt" will possess the desired pharmacological activity of the parent compound. Generally, G.I. S. Paulekuhn, et al. , "Trends in Active Pharmaceutical Ingredient Salt Selection based on Analysis of the Orange Book Database", J. Am. Med. Chem. , 2007, 50:6665-72, S.M. M. Berge, et al. , "Pharmaceutical Salts", J Pharm Sci. , 1977, 66:1-19 and Handbook of Pharmaceutical Salts, Properties, Selection, and Use, Stahl and Wermuth, Eds. , Wiley-VCH and VHCA, Zurich, 2002. Examples of pharmaceutically acceptable salts are salts that are pharmacologically effective and suitable for contact with patient tissue without undue toxicity, irritation, or allergic reactions. The compounds of formula (I) may have sufficiently acidic groups, sufficiently basic groups or both types of functional groups and thus react with a large number of inorganic or organic bases and inorganic and organic acids. to form pharmaceutically acceptable salts.

The present disclosure also relates to pharmaceutically acceptable prodrugs of the compounds of formula (I) and methods of treatment using such pharmaceutically acceptable prodrugs. The term "prodrug" refers to a precursor of a designated compound that produces that compound in vivo after administration to a subject, e.g., through chemical or physiological processes such as solvolysis or enzymatic cleavage, or under physiological conditions. A prodrug that is brought into the body (e.g., brought into physiological pH) is converted to a compound of formula (I). A "pharmaceutically acceptable prodrug" is non-toxic, biologically Specific procedures for selecting and preparing suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed.H. Bundgaard, Elsevier, 1985.

The disclosure also relates to pharmaceutically active metabolites of compounds of formula (I) that may also be used in the disclosed methods. "Pharmaceutically active metabolite" means a pharmacologically active product of metabolism in the body of a compound of formula (I) or a salt thereof. Prodrugs and active metabolites of a compound may be determined using routine techniques known or available in the art. For example, Bertolini, et al. , J Med Chem. 1997, 40, 2011-2016; Shan, et al. , J Pharm Sci. 1997, 86(7), 765-767; Bagshawe, Drug Dev Res. 1995, 34, 220-230; Bodor, Adv Drug Res. １９８４，１３，２２４－３３１；Ｂｕｎｄｇａａｒｄ，Ｄｅｓｉｇｎ ｏｆ Ｐｒｏｄｒｕｇｓ（Ｅｌｓｅｖｉｅｒ Ｐｒｅｓｓ，１９８５）；及びＬａｒｓｅｎ，Ｄｅｓｉｇｎ ａｎｄ Ａｐｐｌｉｃａｔｉｏｎ ｏｆ Ｐｒｏｄｒｕｇｓ，Ｄｒｕｇ Ｄｅｓｉｇｎ ａｎｄ Ｄｅｖｅｌｏｐｍｅｎｔ（Ｋｒｏｇｓｇａａｒｄ－Ｌａｒｓｅｎ，ｅｔ ａｌ．，ｅｄｓ．，Ｈａｒｗｏｏｄ Ａｃａｄｅｍｉｃ Ｐｕｂｌｉｓｈｅｒｓ，１９９１ ).

As used herein, the term "composition" or "pharmaceutical composition" refers to a mixture of at least one compound provided herein and a pharmaceutically acceptable carrier. A pharmaceutical composition facilitates administration of the compound to a patient or subject. There are many techniques of administering compounds in the art, including but not limited to intravenous, oral, aerosol, parenteral, intraocular, intrapulmonary and topical administration.

As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier in or associated with a patient so that the compounds provided herein can perform their intended function. Pharmaceutically acceptable materials such as liquid or solid fillers, stabilizers, dispersants, suspending agents, diluents, excipients, thickeners, solvents or encapsulating materials involved in carrying or transporting , composition or carrier. Typically, such constructs are carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of formulations containing the compounds provided herein and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut, cottonseed, safflower, sesame, olive, corn and soybean; glycols, polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; surfactants; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer; and other non-toxic, compatible substances used in pharmaceutical formulations. As used herein, a "pharmaceutically acceptable carrier" is any coating compatible with the activity of the compounds provided herein and physiologically acceptable to the patient. , antibacterial and antifungal agents and absorption delaying agents and the like. Supplementary active compounds can also be incorporated into the compositions. A "pharmaceutically acceptable carrier" can further include pharmaceutically acceptable salts of the compounds provided herein. Other additional ingredients that can be included in the pharmaceutical compositions provided herein are known in the art, see, for example, Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton , PA). This document is incorporated herein by reference.

The term "stabilizer," as used herein, refers to a polymer capable of chemically inhibiting or preventing the decomposition of compounds of Formula I. Stabilizers can be added to compound formulations to increase the chemical and physical stability of the compounds.

The term "tablet", as used herein, comprises suitable excipients (e.g., fillers, disintegrants, lubricants, glidants and/or surfactants) by conventional tableting processes. shows an orally administrable single-dose solid dosage form that can be prepared by compressing a drug substance, or a pharmaceutically acceptable salt thereof, using . Tablets may be manufactured using conventional granulation methods, such as wet or dry granulation, with optional comminution of granules followed by compression and optional coating. Tablets can also be manufactured by spray drying.

As used herein, the term "capsule" refers to a solid dosage form in which a drug is enclosed within either a hard or soft soluble container or "shell". and/or made of other suitable materials.

As used herein, the terms "effective amount," "pharmaceutically effective amount," and "therapeutically effective amount" refer to a non-toxic but sufficient amount of an agent to provide the desired biological result. refers to the amount of The result may be a reduction or alleviation of the signs, symptoms or causes of disease or any other desired change in a biological system. An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art through routine experimentation.

The terms "combination," "therapeutic combination," "pharmaceutical combination," or "combination product," as used herein, refer to a non-fixed combination or kit-of-parts of combined administration, which In some cases, two or more therapeutic agents can be administered independently at the same time or separately within time intervals, particularly where these time intervals result in the combination partners exhibiting a synergistic effect, e.g., a synergistic effect. becomes possible.

The term "modulators" includes both inhibitors and activators, where "inhibitors" reduce HBV assembly and other HBV core protein functions necessary for HBV replication or generation of infectious particles. , refers to a compound that prevents, inactivates, desensitizes or downregulates.

As used herein, the term "capsid assembly regulator" disrupts or promotes normal capsid assembly (e.g., during maturation) or normal capsid degradation (e.g., during infectivity); or inhibit or interfere with or delay or decrease or modulate or destabilize capsid stability, thereby inducing abnormal capsid morphology and function. In one embodiment, the capsid assembly modifier promotes capsid assembly or disassembly, thereby inducing abnormal capsid morphology. In another embodiment, the capsid assembly regulator interacts with the major capsid assembly protein (CA) (e.g., binds to the active site, binds to the allosteric site, modifies and/or interferes with folding, etc.). ), thereby disrupting capsid assembly or disassembly. In yet another embodiment, capsid assembly regulators destabilize CA structure or function (e.g., CA's ability to assemble, degrade, bind to substrates, fold into a preferred conformation, etc.), weakens viral infectivity and/or kills the virus.

As used herein, the term "treatment" or "treating" means curing, curing, alleviating or reducing HBV infection, symptoms of HBV infection or the likelihood of developing HBV infection. therapeutic agents for patients with HBV infection, symptoms of HBV infection, or potential to develop HBV infection, for the purpose of, altering, correcting, ameliorating, ameliorating or affecting i.e., applying or administering a compound of the disclosure (alone or in combination with another pharmaceutical agent) or therapeutic agents to tissues or cell lines isolated from such patients (e.g., for diagnostic or ex vivo application). defined as the application or administration of Such treatments may be specifically tailored or modified based on knowledge gained from the field of pharmacogenomics.

As used herein, the term "prevent" or "prevention" means no occurrence of a disorder or disease if none had occurred; If it was, it means that no further disorder or disease will develop. The individual's ability to prevent some or all of the symptoms associated with the disorder or disease is also considered.

As used herein, the terms "patient", "individual" or "subject" refer to a human or non-human mammal. Non-human mammals include, for example, farm animals and pets such as sheep, cows, pigs, dogs, cats and rodents. Preferably, the patient, subject or individual is human.

In methods of treatment according to the present disclosure, an effective amount of a pharmaceutical agent according to the present disclosure is administered to a subject suffering from or diagnosed as having such a disease, disorder or condition. . "Effective amount" means an amount or dose sufficient to generally provide the desired therapeutic or prophylactic benefit to a patient in need of such treatment for the specified disease, disorder or condition. The effective amount or dosage of a compound of the present disclosure is determined by routine methods such as, for example, modeling, dose escalation studies or clinical trials, and by routine factors such as mode or route of administration or drug delivery, compound pharmacokinetics, disease, disorder. or the severity and course of the condition, the subject's prior or ongoing therapy, the subject's health status and response to drugs, and the judgment of the treating physician. Exemplary dosages are from about 0.001 to about 200 mg of compound per kg body weight of the subject per day, preferably from about 0.05 to 100 mg/kg/day in single or divided dose units (eg, BID, TID, QID). daily or in the range of about 1-35 mg/kg/day. For a 70 kg human, specific ranges of suitable dosages are about 0.05 to about 7 g/day, or about 0.2 to about 2.5 g/day.

An example dose of the compound is from about 1 mg to about 2,500 mg. In some embodiments, the dose of a compound of the disclosure used in the compositions disclosed herein is less than about 10,000 mg, or less than about 8,000 mg, or less than about 6,000 mg, or less than about 5 mg. ,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, the dose of a second compound described herein (i.e., another drug for treating HBV) is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg; or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg and any whole or partial increments thereof.

Once improvement of the patient's disease, disorder or condition begins, the dose may be adjusted for prophylactic or maintenance therapy. For example, dose or frequency or both can be reduced as a function of symptoms to levels that maintain the desired therapeutic or prophylactic effect. Of course, treatment can be discontinued when symptoms are reduced to an appropriate level. Patients may, however, require intermittent treatment on a long-term basis if symptoms recur.

HBV infections that can be treated by the methods of the present disclosure include HBV genotype A, B, C and/or D infections. However, in one embodiment, the disclosed methods can treat any genotype of HBV (“pan-genotypic therapy”). HBV genotyping can be performed using methods known in the art, eg, INNO-LIPA® HBV Genotyping, Innogenetics N.W. V. , Ghent, Belgium).

Exemplary compounds useful in the methods of the present disclosure will now be described by reference to the specific synthetic schemes below for their general preparation and the specific examples that follow. To obtain the various compounds herein, starting materials are preferred such that ultimately the desired substituents are retained throughout the reaction schemes to give the desired products, with or without protection as appropriate. One of ordinary skill in the art will understand that Alternatively, it may be necessary or desirable to use a suitable group that is retained throughout the reaction scheme and can be optionally substituted with a desired substituent in place of the ultimately desired substituent. Unless otherwise specified, the variables are as defined above with respect to Formula (I). The reaction may be carried out between the melting point and the reflux temperature of the solvent, preferably between 0° C. and the reflux temperature of the solvent. The reaction can be heated using conventional heating or microwave heating. The reaction can also be carried out in a closed pressure vessel at a temperature above the normal reflux temperature of the solvent.

Abbreviations and acronyms used herein include those set forth in Table 2 below.

Synthesis Exemplary compounds useful in the methods of the present disclosure will now be described by reference to the specific synthetic schemes below for their general preparation and the specific examples that follow.

スキーム１によれば、ジクロロメタン（ＤＣＭ）、ＣＣｌ_４などの好適な溶媒中で２－メチリデン－１，３－プロパンジオールを塩化チオニルと反応させて、環状亜硫酸エステルの５－メチレン－１，３，２－ジオキサチアン２－オキシドを得る。この環状亜硫酸を、Ｎ，Ｎ－ジメチルホルムアミド（ＤＭＦ）などの溶媒中、ヒドラジン－１，２－ジカルボン酸ジ－ｔｅｒｔ－ブチルなどの窒素求核試薬、ＮａＨなどの好適な塩基と反応させることにより、１－（２－（ヒドロキシメチル）アリル）ヒドラジン－１，２－ジカルボン酸ジ－ｔｅｒｔ－ブチルが得られる。その後、Ｔ．Ｗ．Ｇｒｅｅｎｅ ａｎｄ Ｐ．Ｇ．Ｍ．Ｗｕｔｓ，“Ｐｒｏｔｅｃｔｉｖｅ Ｇｒｏｕｐｓ ｉｎ Ｏｒｇａｎｉｃ Ｓｙｎｔｈｅｓｉｓ，”３ｅｄ．，Ｊｏｈｎ Ｗｉｌｅｙ＆Ｓｏｎｓ，１９９９に記載されているような確立された方法で脱保護することにより、２－（ヒドラジニルメチル）プロパ－２－エン－１－オールが得られる。

According to Scheme 1, 2-methylidene-1,3-propanediol is reacted with thionyl chloride in a suitable solvent such as dichloromethane (DCM), CCl ₄ to give the cyclic sulfite 5-methylene-1,3, 2-Dioxathiane 2-oxide is obtained. By reacting the cyclic sulfite with a nitrogen nucleophile such as di-tert-butyl hydrazine-1,2-dicarboxylate, a suitable base such as NaH in a solvent such as N,N-dimethylformamide (DMF). , di-tert-butyl 1-(2-(hydroxymethyl)allyl)hydrazine-1,2-dicarboxylate is obtained. After that, T. W. Greene andP. G. M. Wuts, "Protective Groups in Organic Synthesis," 3 ed. , John Wiley & Sons, 1999 to give 2-(hydrazinylmethyl)prop-2-en-1-ol.

スキーム２によれば、式（Ｖ）のオキソピペリジン化合物（式中、Ｒ^４はＨであり、ＰＧはｔｅｒｔ－ブトキシカルボニル保護基（ＢＯＣ基）である）を、ＥｔＯＨなどの好適な溶媒中、２５℃から４０℃の範囲の温度で、約２～５ｈにわたり２－（ヒドロキシメチル）プロパ－２－エン－１－オール、酢酸ナトリウム塩（ＮａＯＡｃ）と縮合させて、式（ＶＩ）の化合物を得る。式（ＶＩ）の化合物を、ＤＭＦなどの好適な溶媒中、４－メチルベンゼンスルホン酸アリル、Ｋ_２ＣＯ_３などの好適な塩基でアルキル化して式（ＶＩＩ）の化合物を得る。式（ＶＩＩ）の化合物の閉環メタセシス反応は、ＤＣＭなどの溶媒中、１６～２４ｈにわたりジクロロ［１，３－ビス（２，４，６－トリメチルフェニル）－２－イミダゾリジニリデン］（２－イソプロポキシフェニルメチレン）ルテニウム（ＩＩ）（Ｈｏｖｅｙｄａ－Ｇｒｕｂｂｓ ＩＩ触媒）を用いて行われ、式（ＶＩＩＩ）の化合物が得られる。式（ＶＩＩＩ）の化合物（式中、Ｒ^４はＣ_１～４アルキルである）は、式（Ｖ）の化合物（式中、Ｒ^４はＣ_１～４アルキルである）から、前述した方法を用いて調製され得る。

According to Scheme 2, an oxopiperidine compound of formula (V), where R ⁴ is H and PG is a tert-butoxycarbonyl protecting group (BOC group), in a suitable solvent such as EtOH, Condensation with 2-(hydroxymethyl)prop-2-en-1-ol, acetic acid sodium salt (NaOAc) at a temperature ranging from 25° C. to 40° C. over a period of about 2-5 h to give a compound of formula (VI). obtain. Compounds of formula (VI) are alkylated with a suitable base such as allyl 4-methylbenzenesulfonate, K ₂ CO ₃ in a suitable solvent such as DMF to give compounds of formula (VII). The ring-closing metathesis reaction of compounds of formula (VII) yields dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](2-iso propoxyphenylmethylene)ruthenium(II) (Hoveyda-Grubbs II catalyst) to give compounds of formula (VIII). A compound of formula (VIII), wherein R ⁴ is C _1-4 alkyl, can be obtained from a compound of formula (V) (wherein R ⁴ is C _1-4 alkyl) by the methods previously described. can be prepared using

スキーム３によれば、式（ＩＸ）の化合物（式中、Ｒ^４はＨであり、ＰＧはＢＯＣである）をトリチオ炭酸ジメチル、ＮａＨなどの塩基と、ＤＭＦなどの溶媒中、０℃～２５℃の範囲の温度で１～３ｈにわたり反応させて、式（Ｘ）の化合物を得る。式（Ｘ）の化合物を、ＥｔＯＨなどの好適な溶媒中、０℃～２５℃の範囲の温度で１２～１６ｈにわたり様々なヒドラジンと縮合させて式（ＸＩ）の化合物を得る。式（ＸＩ）の化合物を（Ｚ）－１，４－ジクロロブタ－２－エン、Ｋ_２ＣＯ_３などの塩基と、ＤＭＦなどの好適な溶媒中、０℃～５０℃の範囲の温度で４～６ｈにわたり反応させて、式（ＸＩＩ）の化合物を得る。式（ＸＩＩ）のアルケン化合物は、テトラヒドロフラン（ＴＨＦ）などの好適な溶媒中、０℃～２５℃の範囲の温度で、ジメチルスルフィドボランによりヒドロホウ素化され、続いて過ホウ酸ナトリウム四水和物により酸化され、式（ＸＩＩＩａ）及び（ＸＩＩＩｂ）の化合物の混合物が得られる。式（ＸＩＩＩａ）及び（ＸＩＩＩｂ）の化合物（式中、Ｒ^４はＣ_１～４アルキルである）は、式（ＩＸ）の化合物（式中、Ｒ^４はＣ_１～４アルキルである）から、前述した方法を用いて調製され得る。

According to Scheme 3, a compound of formula (IX) (wherein R ⁴ is H and PG is BOC) is treated with a base such as dimethyl trithiocarbonate, NaH in a solvent such as DMF from 0° C. to 25° C. C. for 1-3 h to give compounds of formula (X). Compounds of formula (X) are condensed with various hydrazines in a suitable solvent such as EtOH at temperatures ranging from 0° C. to 25° C. for 12-16 h to give compounds of formula (XI). A compound of formula (XI) is treated with a base such as (Z)-1,4-dichlorobut-2-ene, K ₂ CO ₃ in a suitable solvent such as DMF at a temperature ranging from 0°C to 50°C. React for ~6 h to give compounds of formula (XII). Alkene compounds of formula (XII) are hydroborated with dimethylsulfide borane in a suitable solvent such as tetrahydrofuran (THF) at temperatures ranging from 0°C to 25°C followed by sodium perborate tetrahydrate. to give a mixture of compounds of formulas (XIIIa) and (XIIIb). Compounds of formulas (XIIIa) and (XIIIb), wherein R ⁴ is C _1-4 alkyl, are prepared from compounds of formula (IX), wherein R ⁴ is C _1-4 alkyl, to It can be prepared using the methods described above.

スキーム４によれば、式（ＶＩＩＩ）の化合物（式中、Ｒ^４はＨであり、ＰＧはＢＯＣである）を当業者に知られた条件で脱保護して、式（ＸＩＸ）の化合物を得る。続いて、市販の又は合成により入手可能な式（ＸＸ）の化合物（式中、Ｘ、Ｒ^２及びＲ^３は請求項１に定義された通りである）、ＴＥＡなどの好適な塩基と、ＤＣＭなどの好適な溶媒中で反応させることにより、式（Ｉ）の化合物（式中、

は二重結合であり、ＸはＯであり、Ｒ^１はＣＨ_２ＯＨであり、Ｒ^４はＨである）が得られる。式（Ｉ）の化合物（式中、Ｒ^４はＣ_１～４アルキルである）は、式（ＶＩＩＩ）の化合物（式中、Ｒ^４はＣ_１～４アルキルである）から、前述した方法を用いて調製され得る。

According to Scheme 4, compounds of formula (VIII) (wherein ^R4 is H and PG is BOC) are deprotected under conditions known to those skilled in the art to give compounds of formula (XIX). obtain. Subsequently, a commercially available or synthetically available compound of formula (XX), wherein X, ^R2 and R3 ^are as defined in claim 1, a suitable base such as TEA and DCM. Compounds of formula (I) (wherein

is a double bond, X is O, R ¹ is CH ₂ OH and R ⁴ is H). Compounds of formula (I) wherein R ⁴ is C _1-4 alkyl can be obtained from compounds of formula (VIII) (wherein R ⁴ is C _1-4 alkyl) by the methods previously described. can be prepared using

スキーム５によれば、式（ＸＸＩ）の化合物（式中、Ｒ^４はＨであり、ＰＧはＢＯＣである）を当業者に知られた条件で脱保護して、式（ＸＸＩＩ）の化合物を得る。続いて、市販の又は合成により入手可能な式（ＸＸ）の化合物（式中、Ｘ、Ｒ^２及びＲ^３は請求項１に定義された通りである）、ＴＥＡなどの好適な塩基と、ＤＣＭなどの好適な溶媒中で反応させることにより、式（Ｉ）の化合物（式中、

は単結合であり、ＸはＳであり、Ｒ^１はＯＨであり、Ｒ^４はＨである）が得られる。式（Ｉ）の化合物（式中、Ｒ^４はＣ_１～４アルキルである）は、式（ＸＸＩ）の化合物（式中、Ｒ^４はＣ_１～４アルキルである）から、前述した方法を用いて調製され得る。式（Ｉ）の化合物（式中、ＸはＳである）は、当業者に知られた条件を用いて、例えば、ＤＣＭなどの好適な溶媒中、ｍ－ＣＰＢＡ（メタ－クロロ過安息香酸）などの酸化剤を用いて酸化され、式（Ｉ）の化合物（式中、ＸはＳ＝Ｏ又はＳＯ_２であるが得られる）。

According to Scheme 5, compounds of formula (XXI) (wherein ^R4 is H and PG is BOC) are deprotected under conditions known to those skilled in the art to give compounds of formula (XXII). obtain. Subsequently, a commercially available or synthetically available compound of formula (XX), wherein X, ^R2 and R3 ^are as defined in claim 1, a suitable base such as TEA and DCM. Compounds of formula (I) (wherein

is a single bond, X is S, ^R1 is OH and ^R4 is H). Compounds of formula (I) wherein R ⁴ is C _1-4 alkyl can be obtained from compounds of formula (XXI) (wherein R ⁴ is C _1-4 alkyl) by the methods previously described. can be prepared using Compounds of formula (I), wherein X is S, are prepared using conditions known to those skilled in the art, for example m-CPBA (meta-chloroperbenzoic acid) in a suitable solvent such as DCM. to obtain a compound of formula (I) where X is S=O or SO ₂ .

Compounds of formula (I) can be converted to their corresponding salts using methods known to those skilled in the art. For example, amines of formula (I) are treated with trifluoroacetic _acid , HCl or citric _acid in solvents such as _Et2O , CH2Cl2, THF, MeOH, chloroform or isopropanol to give the corresponding salt forms. . Alternatively, salts of trifluoroacetic acid or formic acid are obtained as a result of reverse phase HPLC purification conditions. The crystalline forms of the pharmaceutically acceptable salts of the compounds of formula (I) may be dissolved in polar solvents (including mixtures of polar solvents and aqueous mixtures of polar solvents) or non-polar solvents (including mixtures of non-polar solvents). It can be obtained in crystalline form from crystallization.

Where the compounds according to this disclosure have at least one chiral center, they may therefore exist as enantiomers. Where the compounds possess two or more chiral centers, they may additionally exist as diastereomers. It is understood that all such isomers and mixtures thereof are included within the scope of this disclosure.

Compounds of the formulas represented in the schemes above, represented as "stereoisomeric mixtures" (meaning mixtures of two or more stereoisomers, including enantiomers, diastereomers and combinations thereof) are subject to SFC resolution separated by

Compounds prepared by the above schemes may be obtained as single forms, such as single enantiomers, by form-specific synthesis or resolution. Alternatively, the compounds prepared by the schemes above may be obtained as mixtures of various forms, such as racemic (1:1) mixtures or non-racemic (not 1:1) mixtures. When racemic and non-racemic mixtures of enantiomers are obtained, single enantiomers can be separated by chiral chromatography, recrystallization, diastereomeric salt formation, derivatization to diastereomeric adducts, biotransformation or enzymatic conversion. can be isolated using conventional separation methods known to those skilled in the art, such as Where regioisomeric mixtures or diastereomeric mixtures are obtained, where applicable single isomers may be separated using conventional methods such as chromatography or crystallization.

General Procedures The following specific examples are provided to further illustrate the present disclosure and various preferred embodiments.

In obtaining the compounds and corresponding analytical data described in the examples below, the following experimental and analytical protocols were followed unless otherwise indicated.

Unless otherwise indicated, reaction mixtures were magnetically stirred at room temperature (rt) under a nitrogen atmosphere. Where solutions _were "dried" they _were generally dried with a drying agent such as _Na2SO4 or MgSO4. When mixtures, solutions and extracts are "concentrated," they are typically concentrated using a rotary evaporator under reduced pressure.

Normal phase silica gel chromatography (FCC) was performed on silica gel ( _SiO2 ) using prepacked cartridges.

Preparative reversed phase high performance liquid chromatography (RP HPLC) was performed by either:
Method A. 25 mL/min with Phenomenex Synergi C18 (10 μm, 150×25 mm) or Boston Green ODS C18 (5 μm, 150×30 mm) and mobile phase of 5-99% ACN in water (containing 0.225% FA) for 10 min and then hold at 100% ACN for 2 min, or Method B. Phenomenex Synergi C18 (10 μm, 150×25 mm) or Boston Green ODS C18 (5 μm, 150×30 mm) with a mobile phase of 5-99% ACN in water (0.1% TFA) with a flow rate of 25 mL/min. Gilson GX-281 semi-preparative HPLC or Method C. for 10 minutes at 100% ACN and then 2 min hold at 100% ACN. Phenomenex Synergi C18 (10 μm, 150×25 mm) or Boston Green ODS C18 (5 μm, 150×30 mm) and a mobile phase of 5-99% ACN in water (0.05% HCl) at a flow rate of 25 mL/min Gilson GX-281 semi-preparative HPLC or Method D. for 10 minutes at 100% ACN and then 2 minutes hold at 100% ACN. Phenomenex Gemini C18 (10 μm, 150×25 mm), AD (10 μm, 250×30 mm) or Waters XBridge C18 column (5 μm, 150×30 mm) and 0- in water (containing 0.05 v/v % ammonia hydroxide) Gilson GX-281 semi-preparative HPLC with a mobile phase of 99% ACN with a flow rate of 25 mL/min for 10 minutes and then a 2 min hold at 100% ACN, or Method E. Equipped with a Phenomenex Gemini C18 (10 μm, 150×25 mm) or Waters XBridge C18 column (5 μm, 150×30 mm) and a mobile phase of 5-99% ACN in water (10 mM NH4HCO3) at a flow rate of 25 mL/min for 10 min. Gilson GX-281 semi-preparative HPLC with 2 min hold at 100% ACN.

Preparative supercritical high performance fluid chromatography (SFC) was performed on a Thar 80 Prep-SFC system or a Waters 80Q Prep-SFC system from Waters. The ABPR is set at 100 bar to keep the CO ₂ in SF conditions and the flow rate can be verified according to compound characteristics using flow rates between 50 g/min and 70 g/min. Column temperature was ambient temperature.

Mass spectra (MS) were obtained in positive mode using electrospray ionization (ESI) on a SHIMADZU LCMS-2020 MSD or Agilent 1200\G6110A MSD unless otherwise specified. The calculated (calcd.) mass corresponds to the exact mass.

Nuclear magnetic resonance (NMR) spectra were obtained on a Bruker AVIII 400 spectrometer. The definitions for multiplicity are as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad. For compounds containing exchangeable protons, said protons may or may not be visible on the NMR spectrum, depending on the choice of solvent used to run the NMR spectrum and the concentration of the compound in solution. It will be understood.

Chemical names were generated using ChemDraw Ultra 12.0, ChemDraw Ultra 14.0 (CambridgeSoft Corp., Cambridge, MA) or ACD/Name Version 10.01 (Advanced Chemistry).

Compounds designated as R* or S* are enantiopure compounds whose absolute configuration has not been determined.

工程Ａ．５－メチレン－１，３，２－ジオキサチアン２－オキシド。２－メチレンプロパン－１，３－ジオール（２５．００ｇ、２８３．７７ｍｍｏｌ、２３．１５ｍＬ）のＤＣＭ（１５０．００ｍＬ）溶液にＳＯＣｌ_２（４０．５１ｇ、３４０．５２ｍｍｏｌ、２４．７０ｍＬ）のＤＣＭ（７５．００ｍＬ）溶液を、Ｎ_２下、０℃で加えた。この混合物を０℃で４５ｍｉｎにわたり撹拌した。この混合物を真空下、１５℃で濃縮して、標題化合物（３９．００ｇ、粗製）を黄色油状物として得た。^１ＨＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ＝５．３６（ｄ，Ｊ＝１２．９６Ｈｚ，２Ｈ），５．１５（ｓ，２Ｈ），４．２５（ｄ，Ｊ＝１３．２０Ｈｚ，２Ｈ）。 Intermediate 1: 4-(hydroxymethyl)-5,8,9,11-tetrahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]oxazepine-10 ( 2H)-tert-butyl carboxylate.

Process A. 5-methylene-1,3,2-dioxathiane 2-oxide. To a solution of 2-methylenepropane-1,3-diol (25.00 g, 283.77 mmol, 23.15 mL) in DCM (150.00 mL) was added SOCl ₂ (40.51 g, 340.52 mmol, 24.70 mL) in DCM ( 75.00 mL) solution was added at 0° C. under N ₂ . The mixture was stirred at 0° C. for 45 min. The mixture was concentrated under vacuum at 15° C. to give the title compound (39.00 g, crude) as a yellow oil. < ¹ >H NMR (400 MHz, _CDCl3 ) [delta] = 5.36 (d, J = 12.96 Hz, 2H), 5.15 (s, 2H), 4.25 (d, J = 13.20 Hz, 2H).

Process B. Di-tert-butyl 1-(2-(hydroxymethyl)allyl)hydrazine-1,2-dicarboxylate. NaH (6.56 g, 164.00 mmol, purity 60%) was added to a solution of di-tert-butyl hydrazine-1,2-dicarboxylate (38.09 g, 164.00 mmol, 36.63 mL) in DMF (400.00 mL). It was added in portions at -10°C. After stirring the reaction mixture for 1 h, 5-methylene-1,3,2-dioxathiane 2-oxide (11.00 g, 82.00 mmol) dissolved in DMF (100.00 mL) was added. The mixture was stirred at 60° C. for 20 h. The mixture was poured into HCl (0.5N, 2000 mL) and extracted with ethyl acetate (1500 mL x 2). The combined organic layers were washed with brine (1 L x ₂ ), dried over _Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=10/1 to 3/1) to give the title compound. ¹ H NMR (400 MHz, CDCl ₃ ) δ=6.37 (br s, 1H), 5.14 (br s, 1H), 5.03 (s, 1H), 4.15 (br s, 2H), 4.10 (br s, 2H), 1.47 (s, 18H).

Process C. 2-(Hydroxymethyl)prop-2-en-1-ol. To a solution of di-tert-butyl 1-(2-(hydroxymethyl)allyl)hydrazine-1,2-dicarboxylate (1.10 g, 3.64 mmol) in DCM (10.00 mL) was added trifluoroacetic acid (TFA) (8 .00 mL) was added. The mixture was stirred at 25° C. for 3 hours. The mixture was concentrated in vacuo to give the title compound (1.30g crude, 2TFA) as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 5.38 (s, 1H), 5.25 (s, 1H), 4.14 (s, 2H), 3.67 (s, 2H)

Process D. tert-butyl 3-hydroxy-2-(2-(hydroxymethyl)allyl)-6,7-dihydro-2H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate. To a solution of 2-(hydrazinomethyl)prop-2-en-1-ol (1.22 g, 2TFA) and NaOAc (908.07 mg, 11.07 mmol) in EtOH (3.00 mL) was added 4-oxopiperidine-1, 1-tert-butyl 3-ethyl 3-dicarboxylate (1.00 g, 3.69 mmol) was added. The mixture was stirred at 25° C. for 2 hours. The mixture was concentrated in vacuo. The residue was purified by column chromatography (SiO ₂ , DCM:MeOH=50:1-10:1) to give the title compound (740.00 mg, yield 60.61%) as a yellow solid. MS ₍ ESI): mass calculated for _{C15H23N3O4 309.2} ; m/z found _310.3 [M+H] ^<+ >. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 5.50 (s, 1), 5.23 (s, 1H), 4.49 (s, 2H), 4.25 (S, 2H), 4.10 (s, 2H), 3.70 (t, J=6.0 Hz, 2H), 2.60 (t, J=6.0 Hz, 2H).

Process E. tert-Butyl 3-(allyloxy)-2-(2-(hydroxymethyl)allyl)-6,7-dihydro-2H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate. tert-butyl 3-hydroxy-2-(2-(hydroxymethyl)allyl)-6,7-dihydro-2H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate (320.00 mg, 1 K ₂ CO ₃ (170.83 mg, 1.24 mmol, 1.20 eq) and allyl 4-methylbenzenesulfonate (218.64 mg, 1.03 mmol) in DMF (10.00 mL). ) was added. The mixture was stirred at 15° C. for 19 hrs. The mixture was poured into water (10 mL) and then extracted with ethyl acetate (10 mL x 2). The organic layer was washed with brine ( ₁₀ mL), dried over anhydrous _Na2SO4 and concentrated in vacuo. The residue was purified by RP HPLC (conditions A) to give the title compound (73.00 mg, 192.20 μmol) as a colorless oil. MS ₍ ESI): mass calculated for _{C18H27N3O4 349.2} ; m/z found _350.3 [M+H] ^<+ >.

Process F. 4-(hydroxymethyl)-5,8,9,11-tetrahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]oxazepine-10(2H)-carvone tert-butyl acid. tert-butyl 3-allyloxy-2-[2-(hydroxymethyl)allyl]-6,7-dihydro-4H-pyrazolo[4,3-c]pyridine-5-carboxylate (75.00 mg, 214.64 μmol, 1.00 eq) of dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](2-isopropoxyphenylmethylene)ruthenium(II) in DCM (110.00 mL) (Hoveyda-Grubbs II catalyst) (26.90 mg, 42.93 μmol, 0.20 eq) was added. The mixture was stirred at 15° C. for 16 hr. The mixture was heated to 30° C. and stirred at 30° C. for 16 hr. The mixture was concentrated in vacuo. The residue was purified by column chromatography (SiO ₂ , DCM:MeOH=50:1 to 20:1) to give the title compound (28.50 mg, 41.32% yield) as a colorless oil. MS ₍ ESI): mass calculated for _{C16H23N3O4 321.2} ; m/z found _322.2 [M+H] ^<+ >. ¹ H NMR (400 MHz, CHCl ₃ ) δ = 5.72 (br s, 1H), 4.77 (s, 2H), 4.64 (br s, 2H), 4.34 (br s, 2H), 4.11-4.22 (m, 2H), 3.65 (br s, 2H), 2.66 (br t, J=5.38Hz, 2H), 2.46-2.51 (m, 1H ), 1.48(s, 9H).

工程Ａ．３－（（メチルチオ）カルボノチオイル）－４－オキソピペリジン－１－カルボン酸ｔｅｒｔ－ブチル。４－オキソピペリジン－１－カルボン酸ｔｅｒｔ－ブチル（１０ｇ、５０．１９ｍｍｏｌ）のＤＭＦ（１００ｍＬ）溶液に、Ｎ_２下、０℃で、ＮａＨ（２．６１ｇ、６５．２５ｍｍｏｌ、純度６０％）を添加した。この混合物を０℃で０．５ｈにわたり撹拌した。次いで、トリチオ炭酸ジメチル（９．０２ｇ、６５．２５ｍｍｏｌ）のＤＭＦ（５０ｍＬ）溶液を０℃で加えた。この混合物を２５℃で１ｈにわたり撹拌した。この混合物に飽和ａｑ．ＮＨ_４Ｃｌ（２００ｍＬ）を添加して反応を停止させ、次いで、酢酸エチル（ＥｔＯＡｃ）（６００ｍＬ）で抽出した。有機相を塩水（３００ｍＬ×３）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、真空中で濃縮して、標題化合物（１５．５ｇ、粗製）を黄色油状物として得、これを次の工程で直接使用した。 Intermediate 2: 4-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]thiazepine- tert-butyl 10(11H)-carboxylate.

Process A. tert-Butyl 3-((methylthio)carbonothioyl)-4-oxopiperidine-1-carboxylate. NaH (2.61 g, 65.25 mmol, 60% purity) was added to a solution of tert-butyl 4-oxopiperidine-1-carboxylate (10 g, 50.19 mmol) in DMF (100 mL) under N ₂ at 0 °C. added. The mixture was stirred at 0° C. for 0.5 h. A solution of dimethyl trithiocarbonate (9.02 g, 65.25 mmol) in DMF (50 mL) was then added at 0°C. The mixture was stirred at 25° C. for 1 h. Saturated aq. The reaction was quenched by adding NH ₄ Cl (200 mL) and then extracted with ethyl acetate (EtOAc) (600 mL). The organic phase was washed with brine (300 mL x ₃ ), dried over _Na2SO4 , filtered and concentrated in vacuo to give the title compound (15.5 g, crude) as a yellow oil, which was used directly in the process of

Process B. tert-butyl 3-mercapto-6,7-dihydro-2H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate. To a solution of tert-butyl 3-((methylthio)carbonothioyl)-4-oxopiperidine-1-carboxylate (15.5 g, crude) in EtOH (200 mL) was added N2H4.H2O (2.56 g, 50.21 mmol, 2 .49 mL) was added. The mixture was stirred at 25° C. for 12 h. The reaction mixture was quenched by adding 0.5N HCl (200 mL) at 0° C. and then extracted with EtOAc (400 mL×2). The combined organic layers were washed with brine (600 mL), dried over Na ₂ SO ₄ , filtered, then concentrated under reduced pressure to give the title compound (14.5 g, crude) as a yellow solid, which was Used directly in the next step.

Process C. tert-butyl 5,8,9,11-tetrahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]thiazepine-10(2H)-carboxylate. To a solution of tert-butyl 3-mercapto-6,7-dihydro-2H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate (14.5 g, crude) in DMF (300 mL) was added (Z)- 1,4-Dichlorobut-2-ene (6.90 g, 55.23 mmol) and K ₂ CO ₃ (27.76 g, 200.83 mmol, 4 eq) were added. The mixture was stirred at 50° C. for 4 h. The reaction mixture was quenched with 1N HCl (500 mL) at 0° C. and then extracted with EtOAc (400 mL×2). The combined organic layers were washed with brine ₍ 500 mL), dried over _Na2SO4 , filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO ₂ , petroleum ether/ethyl acetate=3/1 plate 1) to give the title compound (0.8 g, 80% purity) as a yellow oil. MS ₍ ESI): mass calculated for _{C15H21N3O2S 307.1} ; m/z found _308.2 [M+H] ^<+ >. ¹ H NMR (400 MHz, CDCl ₃ ) δ=5.89-5.78 (m, 2H), 4.99 (s, 2H), 4.43 (s, 2H), 3.74-3.64 ( m, 2H), 3.34 (s, 2H), 2.72 (t, J=5.6Hz, 2H), 1.49 (s, 9H).

Process D. 4-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]thiazepine-10 (11H) - tert-butyl carboxylate. of tert-butyl 5,8,9,11-tetrahydro-2H-pyrido[2,3]pyrazolo[2,4-b][1,3]thiazepine-10-carboxylate (0.8 g, 2.08 mmol) BH ₃ .Me ₂ S (10 M, 832.76 μL) was added to a THF (8 mL) solution at 0° C. and the mixture was stirred at 25° C. for 1 h. Sodium perborate tetrahydrate (3.20 g, 20.82 mmol, 4.00 mL) dissolved in _H2O (8 mL) was added at 0 <0>C. The mixture was stirred at 25° C. for 16 h. LCMS showed 35% desired mass and 45% starting material mass detected. The mixture was diluted with EtOAc (40 mL) and washed with brine (40 mL). The organic phase was dried over _Na2SO4 , _filtered and concentrated in vacuo. The residue was purified by RP HPLC (conditions A) to give the title compound (0.15 g, 22.14% yield) and 3-hydroxy-2,3,4,5,8,9-hexahydropyrido [4 ',3':3,4]pyrazolo[5,1-b][1,3]thiazepine-10(11H)-tert-butyl carboxylate (0.09 g, 13.28% yield) as a white solid. Obtained. MS ₍ ESI): mass calculated for _{C15H23N3O3S 325.1} ; m/z found _326.2 [M+H] ^<+ >. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 4.40-4.57 (m, 1H), 4.40 (br s, 2H), 4.23-4.29 (m, 2H), 3.67 -3.71 (m, 2H), 2.88-2.91 (m, 1H), 2.70-2.76 (m, 3H), 1.75-2.10 (m, 2H), 1 .50(s, 9H).

標題化合物を中間体２から分取ＨＰＬＣ（条件Ａ）により単離した。ＭＳ（ＥＳＩ）：Ｃ_１５Ｈ_２３Ｎ_３Ｏ_３Ｓについての計算質量値３２５．１；ｍ／ｚの実測値３２６．２［Ｍ＋Ｈ］^＋。^１Ｈ ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ＝４．５４－４．５３（ｍ，２Ｈ），４．４０（ｂｒ ｓ，２Ｈ），４．０７（ｂｒ ｓ，１Ｈ），３．７１－３．６５（ｍ，２Ｈ），２．９１－２．８８（ｍ，１Ｈ），２．７１－２．６０（ｍ，３Ｈ），２．２４－２．１８（ｍ，２Ｈ），１．５０（ｓ，９Ｈ）． Intermediate 3: 3-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]thiazepine- tert-butyl 10(11H)-carboxylate.

The title compound was isolated from intermediate 2 by preparative HPLC (conditions A). MS ₍ ESI): mass calculated for _{C15H23N3O3S 325.1} ; m/z found _326.2 [M+H] ^<+ >. ¹ H NMR (400 MHz, CDCl ₃ ) δ=4.54-4.53 (m, 2H), 4.40 (br s, 2H), 4.07 (br s, 1H), 3.71-3. 65 (m, 2H), 2.91-2.88 (m, 1H), 2.71-2.60 (m, 3H), 2.24-2.18 (m, 2H), 1.50 ( s, 9H).

工程Ａ．（２，５，８，９，１０，１１－ヘキサヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］オキサゼピン－４－イル）メタノール。４－（ヒドロキシメチル）－５，８，９，１１－テトラヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］オキサゼピン－１０（２Ｈ）－カルボン酸ｔｅｒｔ－ブチル（３０．００ｍｇ、９３．３５μｍｏｌ）のＤＣＭ（３．００ｍＬ）溶液にＴＦＡ（３．０８ｇ、２７．０１ｍｍｏｌ）を加えた。この混合物を１５℃で１ｈｒにわたり撹拌した。混合物を真空中で濃縮して、標題化合物（３１ｍｇ、ＴＦＡ）を無色油状物として得、これを生成することなく次の工程で使用した。 Example 1: N-(3-cyano-4-fluorophenyl)-4-(hydroxymethyl)-5,8,9,11-tetrahydropyrido[4′,3′:3,4]pyrazolo[5, 1-b][1,3]oxazepine-10(2H)-carboxamide.

Process A. (2,5,8,9,10,11-hexahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]oxazepin-4-yl)methanol. 4-(hydroxymethyl)-5,8,9,11-tetrahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]oxazepine-10(2H)-carvone TFA (3.08 g, 27.01 mmol) was added to a solution of tert-butyl acid (30.00 mg, 93.35 μmol) in DCM (3.00 mL). The mixture was stirred at 15° C. for 1 hr. The mixture was concentrated in vacuo to give the title compound (31 mg, TFA) as a colorless oil, which was used in the next step without formation.

Process B. N-(3-cyano-4-fluorophenyl)-4-(hydroxymethyl)-5,8,9,11-tetrahydropyrido[4′,3′:3,4]pyrazolo[5,1-b] [1,3]oxazepine-10(2H)-carboxamide. The resulting 2,5,8,9,10,11-hexahydropyrido[2,3]pyrazolo[2,4-b][1,3]oxazepin-4-ylmethanol (31.00 mg, TFA) and N-(3-cyano-4-fluoro-phenyl)phenylcarbamate (23.69 mg, 92.46 μmol) in DCM (5.00 mL) was added with triethylamine (TEA) (28.07 mg, 277.38 μmol). did. The mixture was stirred at 15° C. for 16 hr. The mixture was concentrated in vacuo. The resulting residue was purified by RP (reverse phase) HPLC (conditions A) and then by RP HPLC (conditions E) to give the title compound (10 mg, 99% pure) as a white solid. MS (ESI): mass calculated for _{C18H18ClFN4O 383.1} ; m/z found _384.1 [M+H] ^<+ >. ¹ H NMR (400 MHz, MeOD) δ 7.82 (dd, J=2.75, 5.56 Hz, 1 H), 7.71 (ddd, J=2.75, 4.71, 9.17 Hz, 1 H) , 7.27 (t, J=8.99 Hz, 1H), 5.80 (br s, 1H), 4.77 (s, 2H), 4.68-4.72 (m, 2H), 4. 48 (s, 2H), 4.09 (s, 2H), 3.80 (t, J=5.81 Hz, 2H), 2.72 (t, J=5.75 Hz, 2H).

標題化合物を、実施例１に類似した方法において、しかし工程Ａで４－ヒドロキシ－２，３，４，５，８，９－ヘキサヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］チアゼピン－１０（１１Ｈ）－カルボン酸ｔｅｒｔ－ブチル（中間体２）を４－（ヒドロキシメチル）－５，８，９，１１－テトラヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］オキサゼピン－１０（２Ｈ）－カルボン酸ｔｅｒｔ－ブチル（中間体１）に代えて用いて調製した。ＭＳ（ＥＳＩ）：Ｃ_１８Ｈ_１８ＦＮ_５Ｏ_２Ｓについての計算質量値３８７．１；ｍ／ｚの実測値３８８．１［Ｍ＋Ｈ］^＋。^１Ｈ ＮＭＲ（４００ＭＨｚ，ＭｅＯＤ）δ＝７．８１（ｄｄ，Ｊ＝２．８，５．６Ｈｚ，１Ｈ），７．７０（ｄｄｄ，Ｊ＝２．８，４．６，９．２Ｈｚ，１Ｈ），７．２７（ｔ，Ｊ＝９．０Ｈｚ，１Ｈ），４．５６（ｓ，１Ｈ），４．５０（ｓ，１Ｈ），４．４４（ｄ，Ｊ＝４．８Ｈｚ，２Ｈ），３．８８（ｂｒ ｓ，１Ｈ），３．８１（ｔ，Ｊ＝５．９Ｈｚ，２Ｈ），２．９３－２．９０（ｍ，１Ｈ），２．７７（ｔ，Ｊ＝５．８Ｈｚ，２Ｈ），２．７３－２．７０（ｍ，１Ｈ），２．３５－２．１３（ｍ，２Ｈ）． Example 2: N-(3-cyano-4-fluorophenyl)-4-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[ 5,1-b][1,3]thiazepine-10(11H)-carboxamide.

The title compound was prepared in a manner analogous to Example 1, but in step A with 4-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[ tert-Butyl 5,1-b][1,3]thiazepine-10(11H)-carboxylate (intermediate 2) was converted to 4-(hydroxymethyl)-5,8,9,11-tetrahydropyrido[4′ ,3′:3,4]pyrazolo[5,1-b][1,3]oxazepine-10(2H)-carboxylate (Intermediate 1) instead of tert-butyl. MS ₍ ESI): mass calculated for _{C18H18FN5O2S 387.1} ; _m /z found 388.1 [M+H] ^<+ >. ¹ H NMR (400 MHz, MeOD) δ = 7.81 (dd, J = 2.8, 5.6 Hz, 1 H), 7.70 (ddd, J = 2.8, 4.6, 9.2 Hz, 1 H ), 7.27 (t, J = 9.0 Hz, 1 H), 4.56 (s, 1 H), 4.50 (s, 1 H), 4.44 (d, J = 4.8 Hz, 2 H), 3.88 (br s, 1H), 3.81 (t, J = 5.9Hz, 2H), 2.93-2.90 (m, 1H), 2.77 (t, J = 5.8Hz, 2H), 2.73-2.70 (m, 1H), 2.35-2.13 (m, 2H).

標題化合物を、実施例１に類似した方法において、しかし工程Ａで３－ヒドロキシ－２，３，４，５，８，９－ヘキサヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］チアゼピン－１０（１１Ｈ）－カルボン酸ｔｅｒｔ－ブチル（中間体３）を４－（ヒドロキシメチル）－５，８，９，１１－テトラヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］オキサゼピン－１０（２Ｈ）－カルボン酸ｔｅｒｔ－ブチル（中間体１）に代えて用いて調製した。ＭＳ（ＥＳＩ）：Ｃ_１８Ｈ_１８ＦＮ_５Ｏ_２Ｓについての計算質量値３８７．１；ｍ／ｚの実測値３８８．１［Ｍ＋Ｈ］^＋。^１Ｈ ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ＝７．７３（ｄｄ，Ｊ＝２．５，５．３Ｈｚ，１Ｈ），７．６７－７．５７（ｍ，１Ｈ），７．１５（ｔ，Ｊ＝８．６Ｈｚ，１Ｈ），６．５６（ｂｒ ｓ，１Ｈ），４．６４－４．５７（ｍ，１Ｈ），４．５０（ｓ，２Ｈ），４．３６－４．２２（ｍ，２Ｈ），３．８１（ｂｒ ｔ，Ｊ＝５．７Ｈｚ，２Ｈ），２．９８－２．５６（ｍ，４Ｈ），２．０７－１．８０（ｍ，２Ｈ）． Example 3: N-(3-cyano-4-fluorophenyl)-3-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[ 5,1-b][1,3]thiazepine-10(11H)-carboxamide.

The title compound was prepared in a manner analogous to Example 1, but in step A with 3-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[ tert-Butyl 5,1-b][1,3]thiazepine-10(11H)-carboxylate (intermediate 3) was converted to 4-(hydroxymethyl)-5,8,9,11-tetrahydropyrido[4′ ,3′:3,4]pyrazolo[5,1-b][1,3]oxazepine-10(2H)-carboxylate (Intermediate 1) instead of tert-butyl. MS ₍ ESI): mass calculated for _{C18H18FN5O2S 387.1} ; _m /z found 388.1 [M+H] ^<+ >. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.73 (dd, J = 2.5, 5.3 Hz, 1H), 7.67-7.57 (m, 1H), 7.15 (t, J =8.6Hz, 1H), 6.56(br s, 1H), 4.64-4.57(m, 1H), 4.50(s, 2H), 4.36-4.22(m, 2H), 3.81 (br t, J=5.7Hz, 2H), 2.98-2.56 (m, 4H), 2.07-1.80 (m, 2H).

Ｎ－（３－シアノ－４－フルオロフェニル）－４－ヒドロキシ－２，３，４，５，８，９－ヘキサヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］チアゼピン－１０（１１Ｈ）－カルボキサミド（８０ｍｇ、２０６．４９μｍｏｌ）のＤＣＭ（３ｍＬ）溶液にｍ－ＣＰＢＡ（５３．４５ｍｇ、２４７．７９μｍｏｌ、純度８０％）を加えた。この混合物を２５℃で１ｈにわたり撹拌した。ＬＣＭＳは、約２９％のスルホキシドと約７０％の出発物質が検出されたことを示した。次いで、別のバッチのｍ－ＣＰＢＡ（１０．６９ｍｇ、４９．５６μｍｏｌ、純度８０％）を加えた。得られた混合物を２５℃でさらに１ｈにわたり撹拌した。ＬＣＭＳは、約１７％のスルホキシド、約４４％のスルホン及び約３１％の出発物質が検出されたことを示した。反応混合物にＮａ_２ＳＯ_３（１０ｍＬ）を０℃で添加することによって反応を停止させ、次いで、ＤＣＭ（５ｍＬ×３）で抽出した。有機層を合わせて塩水（１０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、減圧下で濃縮した。残渣をＲＰ ＨＰＬＣ（条件Ａ）により精製して、標題化合物（５．９５ｍｇ、収率７．００％、純度９８％）及びＮ－（３－シアノ－４－フルオロフェニル）－４－ヒドロキシ－２，３，４，５，８，９－ヘキサヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］チアゼピン－１０（１１Ｈ）－カルボキサミド１、１－ジオキシド（１９．０４ｍｇ、収率２１．７６％、純度９９％）を白色固体として得た。ＭＳ（ＥＳＩ）：Ｃ_１８Ｈ_１８ＦＮ_５Ｏ_３Ｓについての計算質量値４０３．１；ｍ／ｚの実測値４０４．１［Ｍ＋Ｈ］^＋。^１Ｈ ＮＭＲ（４００ＭＨｚ，ＭｅＯＤ）δ＝７．８０（ｄｄｄ，Ｊ＝１．２，２．７，５．６Ｈｚ，１Ｈ），７．６９（ｔｄｄ，Ｊ＝２．４，４．６，９．３Ｈｚ，１Ｈ），７．２７（ｔ，Ｊ＝９．０Ｈｚ，１Ｈ），４．７５－４．５１（ｍ，２Ｈ），４．２０－４．４３（ｍ，１Ｈ），３．８９－３．７９（ｍ，３Ｈ），３．６８－３．３７（ｍ，１Ｈ），３．２６－２．９１（ｍ，１Ｈ），２．８９－２．６２（ｍ，３Ｈ），２．３２－１．９７（ｍ，２Ｈ）． Example 4: N-(3-cyano-4-fluorophenyl)-4-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[ 5,1-b][1,3]thiazepine-10(11H)-carboxamide 1-oxide.

N-(3-cyano-4-fluorophenyl)-4-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[5,1- b][1,3]thiazepine-10(11H)-carboxamide (80 mg, 206.49 μmol) in DCM (3 mL) was added with m-CPBA (53.45 mg, 247.79 μmol, 80% purity). The mixture was stirred at 25° C. for 1 h. LCMS showed ~29% sulfoxide and ~70% starting material detected. Another batch of m-CPBA (10.69 mg, 49.56 μmol, 80% purity) was then added. The resulting mixture was stirred at 25° C. for an additional 1 h. LCMS showed ~17% sulfoxide, ~44% sulfone and ~31% starting material detected. The reaction mixture was quenched by adding Na ₂ SO ₃ (10 mL) at 0° C. and then extracted with DCM (5 mL×3). The combined organic layers were washed with brine ( ₁₀ mL), dried over _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by RP HPLC (conditions A) to give the title compound (5.95 mg, 7.00% yield, 98% purity) and N-(3-cyano-4-fluorophenyl)-4-hydroxy-2 ,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]thiazepine-10(11H)-carboxamide 1,1 -dioxide (19.04 mg, 21.76% yield, 99% purity) was obtained as a white solid. MS ₍ ESI): mass calculated for _{C18H18FN5O3S 403.1} ; m/z found _404.1 [M+H] ^<+ >. ¹ H NMR (400 MHz, MeOD) δ = 7.80 (ddd, J = 1.2, 2.7, 5.6 Hz, 1H), 7.69 (tdd, J = 2.4, 4.6, 9 .3Hz, 1H), 7.27 (t, J = 9.0Hz, 1H), 4.75-4.51 (m, 2H), 4.20-4.43 (m, 1H), 3.89 -3.79 (m, 3H), 3.68-3.37 (m, 1H), 3.26-2.91 (m, 1H), 2.89-2.62 (m, 3H), 2 .32-1.97 (m, 2H).

標題化合物を実施例４から分取ＨＰＬＣ（条件Ａ）により単離した。ＭＳ（ＥＳＩ）：Ｃ_１８Ｈ_１８ＦＮ_５Ｏ_４Ｓについての計算質量値４１９．１；ｍ／ｚの実測値４２０．１［Ｍ＋Ｈ］^＋。^１Ｈ ＮＭＲ（４００ＭＨｚ，ＭｅＯＤ）δ＝７．８０（ｂｒ ｓ，１Ｈ），７．６９（ｂｒ ｓ，１Ｈ），７．２６（ｂｒ ｔ，Ｊ＝８．８Ｈｚ，１Ｈ），４．７７（ｂｒ ｓ，２Ｈ），４．５７（ｂｒ ｄ，Ｊ＝４．６Ｈｚ，３Ｈ），４．０８（ｂｒ ｓ，１Ｈ），３．８１（ｂｒ ｓ，２Ｈ），３．５８（ｂｒ ｔ，Ｊ＝１２．４Ｈｚ，１Ｈ），２．８３（ｂｒ ｓ，２Ｈ），２．３２－２．２１（ｍ，２Ｈ）． Example 5: N-(3-cyano-4-fluorophenyl)-4-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[ 5,1-b][1,3]thiazepine-10(11H)-carboxamide 1,1-dioxide.

The title compound was isolated from Example 4 by preparative HPLC (conditions A). MS ₍ ESI): mass calculated for _{C18H18FN5O4S 419.1} ; m/z found _420.1 [M+H] ^<+ >. ¹ H NMR (400 MHz, MeOD) δ = 7.80 (br s, 1 H), 7.69 (br s, 1 H), 7.26 (br t, J = 8.8 Hz, 1 H), 4.77 ( br s, 2H), 4.57 (br d, J = 4.6 Hz, 3H), 4.08 (br s, 1H), 3.81 (br s, 2H), 3.58 (br t, J = 12.4Hz, 1H), 2.83(br s, 2H), 2.32-2.21(m, 2H).

Ｎ－（３－シアノ－４－フルオロフェニル）－３－ヒドロキシ－２，３，４，５，８，９－ヘキサヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］チアゼピン－１０（１１Ｈ）－カルボキサミド（７０ｍｇ、１８０．６８μｍｏｌ）のＤＣＭ（１．５ｍＬ）溶液にｍ－ＣＰＢＡ（４６．７７ｍｇ、２１６．８１μｍｏｌ、純度８０％）を加え、この混合物を２５℃で１ｈにわたり撹拌した。反応混合物にＮａ_２ＳＯ_３（１０ｍＬ）を０℃で添加することによって反応を停止させ、次いで、ＤＣＭ（５ｍＬ×３）で抽出した。有機層を合わせて塩水（１０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、減圧下で濃縮した。残渣をＲＰ ＨＰＬＣ（条件Ａ）により精製して、標題化合物（１７．３６ｍｇ、収率２３．５８％、純度９９％）及びＮ－（３－シアノ－４－フルオロフェニル）－３－ヒドロキシ－２，３，４，５，８，９－ヘキサヒドロピリド［４’，３’：３，４］ピラゾロ［５，１－ｂ］［１，３］チアゼピン－１０（１１Ｈ）－カルボキサミド１，１－ジオキシド（３１．５８ｍｇ、収率４０．８４％、純度９８％）を白色固体として得た。ＭＳ（ＥＳＩ）：Ｃ_１８Ｈ_１８ＦＮ_５Ｏ_３Ｓについての計算質量値４０３．１；ｍ／ｚの実測値４０４．１［Ｍ＋Ｈ］^＋．^１Ｈ ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ＝７．７３－７．７０（ｍ，１Ｈ），７．６１－７．５８（ｍ，１Ｈ），７．１７－７．１３（ｍ，１Ｈ），６．８２－６．６０（ｍ，１Ｈ），５．３３－５．１０（ｍ，１Ｈ），４．８６－４．４２（ｍ，４Ｈ），４．１８－３．０６（ｍ，１Ｈ），３．９０－３．６８（ｍ，２Ｈ），２．９８－２．６４（ｍ，３Ｈ），２．５６－１．９３（ｍ，２Ｈ）． Example 6: N-(3-cyano-4-fluorophenyl)-3-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[ 5,1-b][1,3]thiazepine-10(11H)-carboxamide 1-oxide.

N-(3-cyano-4-fluorophenyl)-3-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[5,1- b][1,3]thiazepine-10(11H)-carboxamide (70 mg, 180.68 μmol) in DCM (1.5 mL) was added with m-CPBA (46.77 mg, 216.81 μmol, 80% purity), The mixture was stirred at 25° C. for 1 h. The reaction mixture was quenched by adding Na ₂ SO ₃ (10 mL) at 0° C. and then extracted with DCM (5 mL×3). The combined organic layers were washed with brine ( ₁₀ mL), dried over _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by RP HPLC (conditions A) to give the title compound (17.36 mg, 23.58% yield, 99% purity) and N-(3-cyano-4-fluorophenyl)-3-hydroxy-2 ,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[5,1-b][1,3]thiazepine-10(11H)-carboxamide 1,1 -dioxide (31.58 mg, 40.84% yield, 98% purity) was obtained as a white solid. MS ₍ ESI): mass calculated for _{C18H18FN5O3S 403.1} ; m/z found _404.1 [M+H] ^<+ >. ¹ H NMR (400 MHz, CDCl ₃ ) δ=7.73-7.70 (m, 1H), 7.61-7.58 (m, 1H), 7.17-7.13 (m, 1H), 6.82-6.60 (m, 1H), 5.33-5.10 (m, 1H), 4.86-4.42 (m, 4H), 4.18-3.06 (m, 1H) ), 3.90-3.68 (m, 2H), 2.98-2.64 (m, 3H), 2.56-1.93 (m, 2H).

標題化合物を実施例６からＲＰ ＨＰＬＣ（条件Ａ）により単離した。ＭＳ（ＥＳＩ）：Ｃ_１８Ｈ_１８ＦＮ_５Ｏ_４Ｓについての計算質量値４１９．１；ｍ／ｚの実測値４２０．１［Ｍ＋Ｈ］^＋。^１Ｈ ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ＝７．７６（ｄｄ，Ｊ＝２．８，５．４Ｈｚ，１Ｈ），７．６０（ｄｄｄ，Ｊ＝２．９，４．６，９．２Ｈｚ，１Ｈ），７．１５（ｔ，Ｊ＝８．７Ｈｚ，１Ｈ），６．７３（ｓ，１Ｈ），４．８３－４．７０（ｍ，３Ｈ），４．５０－４．４９（ｍ，２Ｈ），３．８４（ｔ，Ｊ＝５．７Ｈｚ，２Ｈ），３．６６－３．４６（ｍ，２Ｈ），２．８７（ｔ，Ｊ＝５．７Ｈｚ，２Ｈ），２．２０（ｂｒ ｓ，２Ｈ）． Example 7: N-(3-cyano-4-fluorophenyl)-3-hydroxy-2,3,4,5,8,9-hexahydropyrido[4′,3′:3,4]pyrazolo[ 5,1-b][1,3]thiazepine-10(11H)-carboxamide 1,1-dioxide.

The title compound was isolated from Example 6 by RP HPLC (conditions A). MS ₍ ESI): mass calculated for _{C18H18FN5O4S 419.1} ; m/z found _420.1 [M+H] ^<+ >. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.76 (dd, J = 2.8, 5.4 Hz, 1 H), 7.60 (ddd, J = 2.9, 4.6, 9.2 Hz, 1H), 7.15 (t, J = 8.7Hz, 1H), 6.73 (s, 1H), 4.83-4.70 (m, 3H), 4.50-4.49 (m, 2H), 3.84 (t, J = 5.7Hz, 2H), 3.66-3.46 (m, 2H), 2.87 (t, J = 5.7Hz, 2H), 2.20 ( br s, 2H).

Biological Data HBV Replication Inhibition Assay Inhibition of HBV replication by the compounds of the present disclosure was measured in cells infected or transfected with HBV, such as HepG2.2.15 cells (Sells et al. 1987), or in cells stably integrated with HBV. measured in 1987). In this example, HepG2.2.15 cells were maintained in cell culture medium containing 10% fetal bovine serum (FBS), Geneticin, L-glutamine, penicillin and streptomycin. HepG2.2.15 cells were seeded in 96-well plates at a density of 40,000 cells/well and serially diluted compounds alone or drugs in a checker box format were added at a final DMSO concentration of 0.5%. were treated with a combination of Cells were incubated with compounds for 3 days after which the medium was removed and fresh medium containing compounds was added to the cells and incubated for an additional 3 days. On day 6, the supernatant was removed and treated with DNase at 37°C for 60 minutes, followed by enzyme inactivation at 75°C for 15 minutes. Encapsidated HBV DNA was released from virions and covalently bound HBV polymerase by incubation in lysis buffer (Affymetrix QS0010) containing 2.5 μg proteinase K at 50° C. for 40 minutes. HBV DNA was denatured by the addition of 0.2 M NaOH and detected using a branched-chain DNA (BDNA) QuantiGene assay kit according to the manufacturer's recommendations (Affymetrix). HBV DNA levels were also measured by amplification of encapsidated HBV DNA extracts and HBV DNA using the QuickExtraction Solution (Epicentre Biotechnologies) using an HBV-specific PCR probe that can hybridize to the HBV DNA and a fluorescently labeled probe for quantification. was quantified using qPCR based on the amplification of Additionally, cell viability of HepG2.2.15 cells incubated with test compounds alone or in combination was determined by using CellTitre-Glo reagent according to the manufacturer's protocol (Promega). The average background signal from wells containing only culture medium was subtracted from all other samples, and the percent inhibition at each compound concentration was calculated using equation E1 for HepG2.2.15 treated with 0.5% DMSO. Calculated by normalizing the signal from the cells.
E1: Inhibition rate (%) = (DMSOave-Xi)/DMSOave x 100%
where DMSOave is the average signal calculated from wells treated with DMSO control (0% inhibition control) and Xi is the signal measured from individual wells. EC50 values, the effective concentration that achieved ₅₀ % inhibitory effect, were determined by non-linear fitting and equation E2 using Graphpad Prism software (San Diego, Calif.).
E2: Y = Ymin + (Ymax - Ymin) / (1 + 10 (LogEC50 - X) x HillSlope)
where Y represents the percent inhibition value and X represents the logarithm of the compound concentration.

Selected disclosed compounds were assayed by the HBV replication assay (BDNA assay) as described above and a representative group of these active compounds are shown in Table 3. Table 3 shows the _EC50 values obtained by the BDNA assay for a group of selected compounds.

The subject matter of this disclosure is not to be limited in scope by the specific embodiments and examples described herein. Indeed, various modifications of the disclosure, in addition to those enumerated herein, will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

All references (e.g., publications or patents or patent applications) cited in this specification are hereby incorporated by reference as if each individual reference (e.g., publications or patents or patent applications) were incorporated in their entirety for any purpose. is hereby incorporated by reference in its entirety to the same extent for all purposes as if specifically and individually indicated to be incorporated by reference. Other embodiments are within the following claims.

Claims (28)

Formula (I):

(In the formula,
R ¹ is selected from the group consisting of F, OH and C _1-6 alkyl;
R ² is selected from the group consisting of Br, CN and C _1-4 haloalkyl;
R ³ is H or F;
R ⁴ is H or C _1-4 alkyl;
X is selected from the group consisting of O, S, S ₌ O and SO2; and Y is selected from the group consisting of CH, CF and N)
and pharmaceutically acceptable salts, solvates, stereoisomers, isotopic variants or N-oxides thereof. 2. The compound of claim 1, wherein ^R1 is OH. 2. The compound of claim 1, wherein ^R1 is F. The compound of claim 1, wherein R ¹ is C _1-6 alkyl. 2. The compound of claim 1, wherein ^R2 is Br, CN or _CF3 . 2. The compound of claim ¹ , wherein R3 is H. 2. The compound of claim ¹ , wherein R3 is F. 2. The compound of claim 1, wherein ^R4 is H. 2. The compound of claim 1, wherein ^R4 is _CH3 . 2. The compound of claim 1, wherein Y is N. 2. The compound of claim 1, wherein Y is CF. 2. The compound of claim 1, wherein Y is CH. 2. The compound of claim 1, wherein X is O. 2. The compound of claim 1, wherein X is S. 2. The compound of claim 1, wherein X is S=O. _2. The compound of claim 1, wherein X is SO2.

is 3-cyano-4-fluorophenyl, 4-fluoro-3-(trifluoromethyl)phenyl, 3-cyano-2,4-difluorophenyl, 3-bromo-2,4-difluorophenyl, 2-(difluoro A compound according to claim 1, which is methyl)-3-fluoropyridin-4-yl or 2-bromo-3-fluoropyridin-4-yl.

The compound of claim 1, wherein is 3-cyano-4-fluorophenyl.

and pharmaceutically acceptable salts, solvates or N-oxides or compounds selected from the group consisting of N-oxides thereof. (A) Formula (I):

(In the formula,
R ¹ is selected from the group consisting of F, OH and C _1-6 alkyl;
R ² is selected from the group consisting of Br, CN and C _1-4 haloalkyl;
R ³ is H or F;
R ⁴ is H or C _1-4 alkyl;
X is selected from the group consisting of O, S, S ₌ O and SO2; and Y is selected from the group consisting of CH, CF and N)
and at least one compound selected from pharmaceutically acceptable salts, solvates, stereoisomers, isotopic variants or N-oxides of compounds of formula (I);
(B) a pharmaceutical composition comprising at least one pharmaceutically acceptable excipient; 20. A pharmaceutical composition comprising at least one compound according to claim 19 and at least one pharmaceutically acceptable excipient. 11. A method of treating HBV infection in an individual in need thereof, comprising administering a therapeutically effective amount of at least one compound according to claim 1 to said individual. 10. A method of inhibiting or reducing the formation or presence of HBV DNA-containing particles or HBV RNA-containing particles in an individual in need thereof, comprising administering a therapeutically effective amount of a compound of claim 1 to said A method comprising administering to an individual. 24. The method of claim 22 or 23, further comprising administering at least one additional therapeutic agent to said individual. Said additional therapeutic agents include HBV polymerase inhibitors, immunomodulatory agents, interferons, viral entry inhibitors, viral maturation inhibitors, capsid assembly modulators, reverse transcriptase inhibitors, cyclophilin/TNF inhibitors, TLR agonists and HBV vaccines 25. The method of claim 24, selected from at least one of the group consisting of: The therapeutic agents include zidovudine, didanosine, zalcitabine, ddA, stavudine, lamivudine, abacavir, emtricitabine, entecavir, apricitabine, atevirapine, ribavirin, acyclovir, famciclovir, valacyclovir, ganciclovir, valganciclovir, tenofovir, adefovir, PMPA, cidofovir, 26. The method of claim 25, wherein the reverse transcriptase inhibitor is selected from the group consisting of efavirenz, nevirapine, delavirdine and etravirine. The therapeutic agents are SM360320 (9-benzyl-8-hydroxy-2-(2-methoxy-ethoxy)-adenine) and AZD8848 ([3-({[3-(6-amino-2-butoxy-8-oxo -7,8-dihydro-9H-purin-9-yl)propyl][3-(4-morpholinyl)propyl]amino}methyl)phenyl]methyl acetate). 25. The method according to 25. 26. The method of claim 25, wherein said therapeutic agent is an HBV vaccine selected from the group consisting of RECOMBIVAX HB, ENGERIX-B, ELOVAC B, GENEVAC-B and SHANVAC B.